bluuebunny/crossref_metadata_2025 huggingface dataset:
Cycling Time Trial Performance 4 Hours After Glycogen-Lowering Exercise Is Similarly Enhanced by Recovery Nondairy Chocolate Beverages Versus Chocolate Milk
Authors: Upshaw Adam U. , Wong Tiffany S. , Bandegan Arash , Lemon Peter W.R.
Published: August 2015

Postexercise chocolate milk ingestion has been shown to enhance both glycogen resynthesis and subsequent exercise performance. To assess whether nondairy chocolate beverage ingestion post–glycogen-lowering exercise can enhance 20-km cycling time trial performance 4 hr later, eight healthy trained male cyclists (21.8 ± 2.3y, VO2max = 61.2 ± 1.4 ml·kg-1·min-1; M ± SD) completed a series of intense cycling intervals designed to lower muscle glycogen (Jentjens & Jeukendrup, 2003) followed by 4 hr of recovery and a subsequent 20-km cycling time trial. During the first 2 hr of recovery, participants ingested chocolate dairy milk (DAIRYCHOC), chocolate soy beverage (SOYCHOC), chocolate hemp beverage (HEMPCHOC), low-fat dairy milk (MILK), or a low-energy artificially sweetened, flavored beverage (PLACEBO) at 30-min intervals in a double-blind, counterbalanced repeated-measures design. All drinks, except the PLACEBO (247 kJ) were isoenergetic (2,107 kJ), and all chocolate-flavored drinks provided 1-g CHO·kg body mass-1·h-1. Fluid intake across treatments was equalized (2,262 ± 148 ml) by ingesting appropriate quantities of water based on drink intake. The CHO:PRO ratio was 4:1, 1.5:1, 4:1, and 6:1 for DAIRYCHOC, MILK, SOYCHOC, and HEMPCHOC, respectively. One-way analysis of variance with repeated measures showed time trial performance (DAIRYCHOC = 34.58 ± 2.5 min, SOYCHOC = 34.83 ± 2.2 min, HEMPCHOC = 34.88 ± 1.1 min, MILK = 34.47 ± 1.7 min) was enhanced similarly vs PLACEBO (37.85 ± 2.1) for all treatments (p = .019) These data suggest that postexercise macronutrient and total energy intake are more important for same-day 20-km cycling time trial performance after glycogen-lowering exercise than protein type or protein-to-carbohydrate ratio.
Gender Differences in Physical Activity Levels of Older People With Type 2 Diabetes Mellitus
Authors: Kelly Joan , Edney Katrina , Moran Chris , Srikanth Velandai , Callisaya Michele
Published: August 2015

Background:

Physical activity (PA) is important in managing Type 2 Diabetes Mellitus (T2DM). This study aimed to determine 1) the number of daily steps taken by older people with T2DM, 2) if T2DM is associated with taking fewer steps per day and less likelihood of meeting PA guidelines, and 3) whether these associations are modified by age or gender.

Methods:

PA was obtained by pedometer from 2 cohorts of older adults with and without T2DM. Multivariable regression was used to determine associations between T2DM, mean steps per day and meeting a guideline equivalent (7 100 steps per day).

Results:

There were 293 participants with T2DM (mean age 67.6 ± 6.8 years) and 336 without T2DM (mean age 72.1 ± 7.1 years). In women, T2DM was associated with fewer mean steps per day (β = –1306.4; 95% CI –2052.5, –560.3; P = .001) and not meeting the PA guidelines (OR 0.51; 95% CI 0.28, 0.92; P = .03). Associations were not significant in men (P > .05). Only 29.7% of those with T2DM and 33.3% of those without T2DM met PA guidelines.

Conclusions:

Greater focus is needed on how to maintain and increase PA in older age with particular focus on women with T2DM.
The Experience of Competition Stress and Emotions in Cricket
Authors: Neil Rich , Bowles Harry C.R. , Fleming Scott , Hanton Sheldon
Published: August 2015

The purpose of the study was to conduct an in-depth examination of the stress and emotion process experienced by three sub-elite-level male cricketers over a series of five competitive performances. Using reflective diaries and follow-up semistructured interviews, the findings highlighted the impact of appraisal, coping, and emotion on performance, with perceptions of control and self-confidence emerging as variables that can influence the emotive and behavioral outcomes of a stressful transaction. Postperformance, guided athlete reflection was advanced as a valuable tool in the production and application of idiographic coping behaviors that could enhance perceptions of control and self-confidence and influence stress and emotion processes.
Psychosocial Development in Youth Soccer Players: Assessing the Effectiveness of the 5Cs Intervention Program
Authors: Harwood Chris G. , Barker Jamie B. , Anderson Richard
Published: August 2015

This study examined the effectiveness of a longitudinal 5C coaching intervention (Harwood, 2008), focused on promoting behavioral responses associated with commitment, communication, concentration, control, and confidence in youth soccer players. Five players, their parents and a youth academy soccer coach participated in a single-case multiple-baseline across individuals design with multiple treatments. Following baseline, the coach received sequential education in the principles of each C subsequent to integrating relevant strategies in their coaching sessions. During the five intervention phases, players completed assessments of their behavior in training associated with each C, triangulated with observationbased assessments by the coach and the players’ parents. Results indicated psychosocial improvements with cumulative increases in positive psychosocial responses across the intervention for selected players. Changes in player behavior were also corroborated by parent and coach data in conjunction with postintervention social validation. Findings are discussed with respect to the processes engaged in the intervention, and the implications for practitioners and applied researchers.
Many psychology papers fail replication test
Authors: Bohannon John
Published: August 2015

An effort to repeat 100 studies yields sobering results, but many researchers are positive about the process.
Orchids' dazzling diversity explained
Authors: Stokstad Erik
Published: August 2015

New family tree shows that a series of innovations accelerated speciation.
Birth of the moralizing gods
Authors: Wade Lizzie
Published: August 2015

A new theory aims to explain the success of world religions—but testing it remains a challenge.
Global competition propels innovation “snowball”
Authors: Jarvis Michaela
Published: August 2015

In Nairobi, GIST Tech-I finalists transformed their ideas into business plans for great social impact
Response to Comment on “Global diversity and geography of soil fungi”
Authors: Tedersoo Leho , Bahram Mohammad , Põlme Sergei , Anslan Sten , Riit Taavi , Kõljalg Urmas , Nilsson R. Henrik , Hildebrand Falk , Abarenkov Kessy
Published: August 2015

Schadt and Rosling (Technical Comment, 26 June 2015, p. 1438) argue that primer-template mismatches neglected the fungal class Archaeorhizomycetes in a global soil survey. Amplicon-based metabarcoding of nine barcode-primer pair combinations and polymerase chain reaction (PCR)–free shotgun metagenomics revealed that barcode and primer choice and PCR bias drive the diversity and composition of microorganisms in general, but the Archaeorhizomycetes were little affected in the global study. We urge that careful choice of DNA markers and primers is essential for ecological studies using high-throughput sequencing for identification.
Global assessment of arbuscular mycorrhizal fungus diversity reveals very low endemism
Authors: Davison J. , Moora M. , Öpik M. , Adholeya A. , Ainsaar L. , Bâ A. , Burla S. , Diedhiou A. G. , Hiiesalu I. , Jairus T. , Johnson N. C. , Kane A. , Koorem K. , Kochar M. , Ndiaye C. , Pärtel M. , Reier Ü. , Saks Ü. , Singh R. , Vasar M. , Zobel M.
Published: August 2015

Cosmopolitan plant root symbionts

The aboveground lives of plants are only sustainable because of the symbiotic soil fungi that encase their roots. These fungi swap nutrients with plants, defend them from attack, and help them withstand abrupt environmental changes. Out of necessity, fungal symbionts in the soil would appear to be restricted and local to certain plant species. Davison
et al.
, however, discovered that some taxa are globally distributed. How these underground fungi have dispersed so widely remains a mystery; perhaps human farmers have had something to do with it.

Science
, this issue p.
970
Defining the genus
Homo
Authors: Schwartz Jeffrey H. , Tattersall Ian
Published: August 2015

Early hominin species were as diverse as other mammals
Iron-catalyzed intermolecular [2+2] cycloadditions of unactivated alkenes
Authors: Hoyt Jordan M. , Schmidt Valerie A. , Tondreau Aaron M. , Chirik Paul J.
Published: August 2015

Iron plays matchmaker to pair up olefins

In theory, shining the right wavelength of light onto carbon-carbon double bonds should pair them up into four-membered cyclobutane rings. In practice, however, this route can prove finicky and inefficient, particularly if the necessary wavelength lies deep in the ultraviolet region. Hoyt
et al.
report an iron catalyst that coaxes a wide variety of simple olefins into such rings without the need for photoexcitation (see the Perspective by Smith and Baran). Systematic optimization of the ligand coordinated to iron effectively eliminated competing pathways to alternative products.

Science
, this issue p.
960
; see also p.
925
The daring doctor
Sonic Wind
The Story of John Paul Stapp and How a Renegade Doctor Became the Fastest Man on Earth
Craig Ryan
Liveright, 2015. 411 pp.
Authors: Vinsel Lee
Published: August 2015

John Paul Stapp and the biophysical boundaries of the human body
Production of amorphous nanoparticles by supersonic spray-drying with a microfluidic nebulator
Authors: Amstad Esther , Gopinadhan Manesh , Holtze Christian , Osuji Chinedum O. , Brenner Michael P. , Spaepen Frans , Weitz David A.
Published: August 2015

Crystal nuclei beaten to the punch

Amorphous nanoparticles often dissolve more rapidly than their crystalline counterparts, which can be useful in applications such as drug delivery. Amstad
et al.
made amorphous nanoparticles from organic and inorganic compounds—even table salt—using droplets of dissolved compounds created with a microfluidic nebulator. The solvent evaporates fast enough that nanoparticles form before crystal nuclei can develop. The small particle size inhibits crystallization for periods of months

Science
, this issue p.
956
As simple as [2+2]
Authors: Smith Myles W. , Baran Phil S.
Published: August 2015

Iron catalysis transforms readily available commodity olefins into cyclobutane building blocks under thermal conditions

[Also see Report by

Hoyt
et al.

]
Microbiota RORgulates intestinal suppressor T cells
Authors: Hegazy Ahmed N. , Powrie Fiona
Published: August 2015

Gut microbes influence the balance of regulatory T cell subtypes to control inflammation

[Also see Reports by

Ohnmacht
et al.

and

Sefik
et al.

]
Synthetic communities, the sum of parts
Authors: Teague Brian P. , Weiss Ron
Published: August 2015

Complex behaviors are engineered from cooperating cell communities

[Also see Report by

Chen
et al.

]
Generation and Efficacy Evaluation of a Recombinant Pseudorabies Virus Variant Expressing the E2 Protein of Classical Swine Fever Virus in Pigs
Authors: Wang Yimin , Yuan Jin , Cong Xin , Qin Hua-Yang , Wang Chun-Hua , Li Yongfeng , Li Su , Luo Yuzi , Sun Yuan , Qiu Hua-Ji
Published: August 2015

Classical swine fever (CSF) is an economically important infectious disease of pigs caused by classical swine fever virus (CSFV). Pseudorabies (PR), which is caused by pseudorabies virus (PRV), is another important infectious disease of pigs and other animals. Coinfections of pigs with PRV and CSFV occur occasionally in the field. The modified live vaccine Bartha-K61 strain has played an important role in the control of PR in many countries, including China. Since late 2011, however, increasing PR outbreaks caused by an emerging PRV variant have been reported in Bartha-K61-vaccinated swine populations on many farms in China. Previously, we generated a gE/gI-deleted PRV (rPRVTJ-delgE) based on this PRV variant, which was shown to be safe and can provide rapid and complete protection against lethal challenge with the PRV variant in pigs. Here, we generated a new recombinant PRV variant expressing the E2 gene of CSFV (rPRVTJ-delgE/gI-E2) and evaluated its immunogenicity and efficacy in pigs. The results showed that rPRVTJ-delgE/gI-E2 was safe for pigs, induced detectable anti-PRV and anti-CSFV neutralizing antibodies, and provided complete protection against the lethal challenge with either the PRV TJ strain or the CSFV Shimen strain. The data indicate that rPRVTJ-delgE/gI-E2 is a promising candidate bivalent vaccine against PRV and CSFV coinfections.
Immunization with Pneumococcal Surface Protein K of Nonencapsulated Streptococcus pneumoniae Provides Protection in a Mouse Model of Colonization
Authors: Keller Lance E. , Luo Xiao , Thornton Justin A. , Seo Keun-Seok , Moon Bo Youn , Robinson D. Ashley , McDaniel Larry S.
Published: August 2015

Current vaccinations are effective against encapsulated strains of
Streptococcus pneumoniae
, but they do not protect against nonencapsulated
Streptococcus pneumoniae
(NESp), which is increasing in colonization and incidence of pneumococcal disease. Vaccination with pneumococcal proteins has been assessed for its ability to protect against pneumococcal disease, but several of these proteins are not expressed by NESp. Pneumococcal surface protein K (PspK), an NESp virulence factor, has not been assessed for immunogenic potential or host modulatory effects. Mammalian cytokine expression was determined in an
in vivo
mouse model and in an
in vitro
cell culture system. Systemic and mucosal mouse immunization studies were performed to determine the immunogenic potential of PspK. Murine serum and saliva were collected to quantitate specific antibody isotype responses and the ability of antibody and various proteins to inhibit epithelial cell adhesion. Host cytokine response was not reduced by PspK. NESp was able to colonize the mouse nasopharynx as effectively as encapsulated pneumococci. Systemic and mucosal immunization provided protection from colonization by PspK-positive (PspK
+
) NESp. Anti-PspK antibodies were recovered from immunized mice and significantly reduced the ability of NESp to adhere to human epithelial cells. A protein-based pneumococcal vaccine is needed to provide broad protection against encapsulated and nonencapsulated pneumococci in an era of increasing antibiotic resistance and vaccine escape mutants. We demonstrate that PspK may serve as an NESp target for next-generation pneumococcal vaccines. Immunization with PspK protected against pneumococcal colonization, which is requisite for pneumococcal disease.
Screening for Intermediately Vancomycin-Susceptible and Vancomycin-Heteroresistant Staphylococcus aureus by Use of Vancomycin-Supplemented Brain Heart Infusion Agar Biplates: Defining Growth Interpretation Criteria Based on Gold Standard Confirmation
Authors: Khatib Riad , Riederer Kathleen , Sharma Mamta , Shemes Stephen , Iyer Sugantha P. , Szpunar Susan
Published: August 2015

BHI agars supplemented with vancomycin 4 (BHI-V4) and 3 (BHI-V3) mg/liter have been proposed for screening vancomycin intermediately susceptible
Staphylococcus aureus
(VISA) and heteroresistant (hVISA) phenotypes, respectively, but growth interpretation criteria have not been established. We reviewed the growth results (CFU) during population analysis profile-area under the curve (PAP-AUC) of consecutive methicillin-resistant
Staphylococcus aureus
(MRSA) blood isolates, which were saved intermittently between 1996 and 2012. CFU counts on BHI-V4 and BHI-V3 plates were stratified according to PAP-AUC interpretive criteria: <0.90 (susceptible [S-MRSA]), 0.90 to 1.3 (hVISA), and >1.3 (VISA). CFU cutoffs that best predict VISA and hVISA were determined with the use of receiver operating characteristic (ROC) curves. Mu3, Mu50, and methicillin-susceptible
S. aureus
(MSSA) controls were included. We also prospectively evaluated manufacturer-made BHI-V3/BHI-V4 biplates for screening of 2010-2012 isolates. The PAP-AUC of 616 clinical samples was consistent with S-MRSA, hVISA, and VISA in 550 (89.3%), 48 (7.8%), and 18 (2.9%) instances, respectively. For VISA screening on BHI-V4, a cutoff of 2 CFU/droplet provided 100% sensitivity and 97.7% specificity. To distinguish VISA from hVISA, a cutoff of 16 CFU provided 83.3% sensitivity and 94.7% specificity; the specificity was lowered to 89.5% with a 12-CFU cutoff. For detecting hVISA/VISA on BHI-V3, a 2-CFU/droplet cutoff provided 98.5% sensitivity and 93.8% specificity. These results suggest that 2-CFU/droplet cutoffs on BHI-V4 and BHI-V3 best approximate VISA and hVISA gold standard confirmation, respectively, with minimal overlap in samples with borderline PAP-AUC. Simultaneous screening for VISA/hVISA on manufacturer-made BHI-V4/BHI-V3 biplates is easy to standardize and may reduce the requirement for PAP-AUC confirmation.
Comparison of the Carba NP, Modified Carba NP, and Updated Rosco Neo-Rapid Carb Kit Tests for Carbapenemase Detection
Authors: AbdelGhani Sameh , Thomson Gina K. , Snyder James W. , Thomson Kenneth S.
Published: August 2015

The accurate detection of carbapenemase-producing organisms is a major challenge for clinical laboratories. The Carba NP test is highly accurate but inconvenient, as it requires frequent preparation of fresh imipenem solution. The current study was designed to compare the Carba NP test to two alternative tests for accuracy and convenience. These were a modified Carba NP test that utilized intravenous (i.v.) imipenem-cilastatin, which is less expensive than reference standard imipenem powder, and an updated version of the Rosco Neo-Rapid Carb kit, which does not require the preparation of imipenem solution and has a shelf life of 2 years. The comparison included 87 isolates that produced class A carbapenemases (including KPC-2, -3, -4, -5, -6, and -8, NMC-A, and SME type), 40 isolates that produced metallo-β-lactamases (including NDM-1, GIM-1, SPM-1, IMP-1, -2, -7, -8, -18, and -27, and VIM-1, -2, and -7), 11 isolates that produced OXA-48, and one isolate that produced OXA-181. Negative controls consisted of 50 isolates that produced extended-spectrum β-lactamases (ESBLs), AmpCs (including hyperproducers), K1, other limited-spectrum β-lactamases, and porin and efflux mutants. Each test exhibited 100% specificity and high sensitivity (Carba NP, 100%; Rosco, 99% using modified interpretation guidelines; and modified Carba NP, 96%). A modified approach to interpretation of the Rosco test was necessary to achieve the sensitivity of 99%. If the accuracy of the modified interpretation is confirmed, the Rosco test is an accurate and more convenient alternative to the Carba NP test.
Genomic Variability of Serial Human Isolates of Salmonella enterica Serovar Typhimurium Associated with Prolonged Carriage
Authors: Octavia Sophie , Wang Qinning , Tanaka Mark M. , Sintchenko Vitali , Lan Ruiting
Published: August 2015

Salmonella enterica
serovar Typhimurium is an important foodborne human pathogen that often causes self-limiting but severe gastroenteritis. Prolonged excretion of
S
. Typhimurium after the infection can lead to secondary transmissions. However, little is known about within-host genomic variation in bacteria associated with asymptomatic shedding. Genomes of 35 longitudinal isolates of
S
. Typhimurium recovered from 11 patients (children and adults) with culture-confirmed gastroenteritis were sequenced. There were three or four isolates obtained from each patient. Single nucleotide polymorphisms (SNPs) were analyzed in these isolates, which were recovered between 1 and 279 days after the initial diagnosis. Limited genomic variation (5 SNPs or fewer) was associated with short- and long-term carriage of
S
. Typhimurium. None of the isolates was shown to be due to reinfection. SNPs occurred randomly, and the majority of the SNPs were nonsynonymous. Two nonsense mutations were observed. A nonsense mutation in
flhC
rendered the isolate nonmotile, whereas the significance of a nonsense mutation in
yihV
is unknown. The estimated mutation rate is 1.49 × 10
−6
substitution per site per year.
S
. Typhimurium isolates excreted in stools following acute gastroenteritis in children and adults demonstrated limited genomic variability over time, regardless of the duration of carriage. These findings have important implications for the detection of possible transmission events suspected by public health genomic surveillance of
S
. Typhimurium infections.
Prevalence, Risk Factors, and Transmission Dynamics of Extended-Spectrum-β-Lactamase-Producing Enterobacteriaceae: a National Survey of Cattle Farms in Israel in 2013
Authors: Adler Amos , Sturlesi Na'ama , Fallach Noga , Zilberman-Barzilai Deniz , Hussein Omar , Blum Shlomo E. , Klement Eyal , Schwaber Mitchell J. , Carmeli Yehuda
Published: August 2015

Our objectives were to study the prevalence, risk factors for carriage, and transmission dynamics of extended-spectrum-β-lactamase (ESBL)-producing
Enterobacteriaceae
(ESBLPE) in a national survey of cattle. This was a point prevalence study conducted from July to October 2013 in Israel. Stool samples were collected from 1,226 cows in 123 sections on 40 farms of all production types. ESBLPE were identified in 291 samples (23.7%): 287 contained
Escherichia coli
and 4 contained
Klebsiella pneumoniae
. The number of ESBLPE-positive cows was the highest in quarantine stations and on fattening farms and was the lowest on pasture farms (
P
= 0.03). The number of ESBLPE-positive cows was the lowest in sections containing adult cows (age, >25 months) and highest in sections containing calves (age, <4 months) (
P
< 0.001). Infrastructure variables that were significant risk factors for ESBLPE carriage included crowding, a lack of manure cleaning, and a lack of a cooling (
P
< 0.001 for each), all of which were more common in sections containing calves. Antimicrobial prophylaxis was given almost exclusively to calves and was associated with a high number of ESBLPE carriers (
P
< 0.001). The 287
E. coli
isolates were typed into 106 repetitive extragenic palindromic (REP)-PCR types and mostly harbored
bla
CTX-M-1
or
bla
CTX-M-9
group genes. The isolates on the six farms with ≥15 isolates of ESBLPE were of 4 to 7 different REP-PCR types, with one dominant type being harbored by about half of the isolates. Fourteen types were identified on more than one farm, with only six of the farms being adjacent to each other. The prevalence of ESBLPE carriage is high in calves in cowsheds where the use of antimicrobial prophylaxis is common. ESBLPE disseminate within cowsheds mainly by clonal spread, with limited intercowshed transmission occurring.
Clinical Characteristics, Laboratory Identification, and
In Vitro
Antifungal Susceptibility of Yarrowia (Candida) lipolytica Isolates Causing Fungemia: a Multicenter, Prospective Surveillance Study
Authors: Zhao Ying , Chan Jasper Fuk-Woo , Tsang Chi-Ching , Wang He , Guo Dawen , Pan Yuhong , Xiao Yuling , Yue Na , Chen Jonathan Hon-Kwan , Lau Susanna Kar-Pui , Xu Yingchun , Woo Patrick Chiu-Yat
Published: August 2015

Our case series showed that uncomplicated
Yarrowia lipolytica
fungemia might be treated with catheter removal alone. The Vitek 2 YST identification (ID) card system, matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS), and internal transcribed spacer and 25S nuclear ribosomal DNA (nrDNA) gene sequencing provided reliable identification. All isolates had low MICs to voriconazole, echinocandins, and amphotericin B.
Multiplex Molecular Panels for Diagnosis of Gastrointestinal Infection: Performance, Result Interpretation, and Cost-Effectiveness
Authors: Binnicker Matthew J.
Published: August 2015

Gastrointestinal disease is a major cause of morbidity and mortality worldwide, especially among young children and immunocompromised patients. Diarrhea may result from infection with a variety of microbial pathogens, including bacteria, viruses, or parasites. Historically, the diagnosis of infectious diarrhea has been made using microscopy, antigen tests, culture, and real-time PCR. A combination of these traditional tests is often required due to the inability to distinguish between infectious etiologies based on the clinical presentation alone. Recently, several multiplex molecular assays have been developed for the detection of gastrointestinal pathogens directly from clinical stool samples. These panels allow for the detection and identification of up to 20 pathogens in as little as 1 h. This review will focus on the multiplex molecular panels that have received clearance from the FDA for the diagnosis of diarrheal disease and will highlight issues related to test performance, result interpretation, and cost-effectiveness of these new molecular diagnostic tools.
Performance of Xpert MTB/RIF Assay in Diagnosis of Pleural Tuberculosis by Use of Pleural Fluid Samples
Authors: Rufai Syed Beenish , Singh Amit , Kumar Parveen , Singh Jitendra , Singh Sarman
Published: August 2015

Prospectively, 162 pleural fluid samples from patients with probable tuberculous pleural effusion were tested by the Xpert MTB/RIF assay and the Bactec MGIT-960 culture system. Of these, 43 (26.5%) were positive in the MGIT-960 culture, and 23 (14.2%), in the Xpert MTB/RIF assay. The sensitivity and specificity of the Xpert MTB/RIF compared with the MGIT-960 culture were 54.8% and 100%, respectively.
Commentary: Fluoroquinolone Resistance in Salmonella and the Utility of Perfloxacin Disk Diffusion
Authors: Fang Ferric C.
Published: August 2015

Fluoroquinolone resistance is a serious and increasingly common problem in
Salmonella
. Two companion studies in this issue of the
Journal of Clinical Microbiology
(E. Deak, R. Skov, J. A. Hindler, and R. M. Humphries, J Clin Microbiol 53:3405–3410, 2015,
http://dx.doi.org/10.1128/JCM.01393-15
; R. Skov, E. Matuschek, M. Sjölund-Karlsson, J. Åhman, A. Petersen, M. Stegger, M. Torpdahl, and G. Kahlmeter, J Clin Microbiol 53:3411–3417, 2015,
http://dx.doi.org/10.1128/JCM.01287-15
) provide data to support the use of perfloxacin disk diffusion as a convenient and inexpensive surrogate laboratory method to detect fluoroquinolone resistance in
Salmonella
when the direct measurement of fluoroquinolone MICs is not feasible. Recently updated CLSI and EUCAST susceptibility breakpoints will help to optimize clinical outcomes and reduce the likelihood of emergent resistance.
Role of the DNA Sensor STING in Protection from Lethal Infection following Corneal and Intracerebral Challenge with Herpes Simplex Virus 1
Authors: Parker Zachary M. , Murphy Aisling A. , Leib David A.
Published: August 2015

STING is a protein in the cytosolic DNA and cyclic dinucleotide sensor pathway that is critical for the initiation of innate responses to infection by various pathogens. Consistent with this, herpes simplex virus 1 (HSV-1) causes invariable and rapid lethality in STING-deficient (STING
−/−
) mice following intravenous (i.v.) infection. In this study, using real-time bioluminescence imaging and virological assays, as expected, we demonstrated that STING
−/−
mice support greater replication and spread in ocular tissues and the nervous system. In contrast, they did not succumb to challenge via the corneal route even with high titers of a virus that was routinely lethal to STING
−/−
mice by the i.v. route. Corneally infected STING
−/−
mice also showed increased periocular disease and increased corneal and trigeminal ganglia titers, although there was no difference in brain titers. They also showed elevated expression of tumor necrosis factor alpha (TNF-α) and CXCL9 relative to control mice but surprisingly modest changes in type I interferon expression. Finally, we also showed that HSV strains lacking the ability to counter autophagy and the PKR-driven antiviral state had near-wild-type virulence following intracerebral infection of STING
−/−
mice. Together, these data show that while STING is an important component of host resistance to HSV in the cornea, its previously shown immutable role in mediating host survival by the i.v. route was not recapitulated following a mucosal infection route. Furthermore, our data are consistent with the idea that HSV counters STING-mediated induction of the antiviral state and autophagy response, both of which are critical factors for survival following direct infection of the nervous system.

IMPORTANCE
HSV infections represent an incurable source of morbidity and mortality in humans and are especially severe in neonatal and immunocompromised populations. A key step in the development of an immune response is the recognition of microbial components within infected cells. The host protein STING is important in this regard for the recognition of HSV DNA and the subsequent triggering of innate responses. STING was previously shown to be essential for protection against lethal challenge from intravenous HSV-1 infection. In this study, we show that the requirement for STING depends on the infection route. In addition, STING is important for appropriate regulation of the inflammatory response in the cornea, and our data are consistent with the idea that HSV modulates STING activity through inhibition of autophagy. Our results elucidate the importance of STING in host protection from HSV-1 and demonstrate the redundancy of host protective mechanisms, especially following mucosal infection.
Out of the Reservoir: Phenotypic and Genotypic Characterization of a Novel Cowpox Virus Isolated from a Common Vole
Authors: Hoffmann Donata , Franke Annika , Jenckel Maria , Tamošiūnaitė Aistė , Schluckebier Julia , Granzow Harald , Hoffmann Bernd , Fischer Stefan , Ulrich Rainer G. , Höper Dirk , Goller Katja , Osterrieder Nikolaus , Beer Martin
Published: August 2015

The incidence of human cowpox virus (CPXV) infections has increased significantly in recent years. Serological surveys have suggested wild rodents as the main CPXV reservoir. We characterized a CPXV isolated during a large-scale screening from a feral common vole. A comparison of the full-length DNA sequence of this CPXV strain with a highly virulent pet rat CPXV isolate showed a sequence identity of 96%, including a large additional open reading frame (ORF) of about 6,000 nucleotides which is absent in the reference CPXV strain Brighton Red. Electron microscopy analysis demonstrated that the vole isolate, in contrast to the rat strain, forms A-type inclusion (ATI) bodies with incorporated virions, consistent with the presence of complete
ati
and
p4c
genes. Experimental infections showed that the vole CPXV strain caused only mild clinical symptoms in its natural host, while all rats developed severe respiratory symptoms followed by a systemic rash. In contrast, common voles infected with a high dose of the rat CPXV showed severe signs of respiratory disease but no skin lesions, whereas infection with a low dose led to virus excretion with only mild clinical signs. We concluded that the common vole is susceptible to infection with different CPXV strains. The spectrum ranges from well-adapted viruses causing limited clinical symptoms to highly virulent strains causing severe respiratory symptoms. In addition, the low pathogenicity of the vole isolate in its eponymous host suggests a role of common voles as a major CPXV reservoir, and future research will focus on the correlation between viral genotype and phenotype/pathotype in accidental and reservoir species.

IMPORTANCE
We report on the first detection and isolation of CPXV from a putative reservoir host, which enables comparative analyses to understand the infection cycle of these zoonotic orthopox viruses and the relevant genes involved.
In vitro
studies, including whole-genome sequencing as well as
in vivo
experiments using the Wistar rat model and the vole reservoir host allowed us to establish links between genomic sequences and the
in vivo
properties (virulence) of the novel vole isolate in comparison to those of a recent zoonotic CPXV isolated from pet rats in 2009. Furthermore, the role of genes present only in a reservoir isolate can now be further analyzed. These studies therefore allow unique insights and conclusions about the role of the rodent reservoir in CPXV epidemiology and transmission and about the zoonotic threat that these viruses represent.
Genome-Wide Screen Reveals Valosin-Containing Protein Requirement for Coronavirus Exit from Endosomes
Authors: Wong Hui Hui , Kumar Pankaj , Tay Felicia Pei Ling , Moreau Dimitri , Liu Ding Xiang , Bard Frédéric
Published: August 2015

Coronaviruses are RNA viruses with a large zoonotic reservoir and propensity for host switching, representing a real threat for public health, as evidenced by severe acute respiratory syndrome (SARS) and the emerging Middle East respiratory syndrome (MERS). Cellular factors required for their replication are poorly understood. Using genome-wide small interfering RNA (siRNA) screening, we identified 83 novel genes supporting infectious bronchitis virus (IBV) replication in human cells. Thirty of these hits can be placed in a network of interactions with viral proteins and are involved in RNA splicing, membrane trafficking, and ubiquitin conjugation. In addition, our screen reveals an unexpected role for valosin-containing protein (VCP/p97) in early steps of infection. Loss of VCP inhibits a previously uncharacterized degradation of the nucleocapsid N protein. This inhibition derives from virus accumulation in early endosomes, suggesting a role for VCP in the maturation of virus-loaded endosomes. The several host factors identified in this study may provide avenues for targeted therapeutics.

IMPORTANCECoronaviruses are RNA viruses representing a real threat for public health, as evidenced by SARS and the emerging MERS. However, cellular factors required for their replication are poorly understood. Using genome-wide siRNA screening, we identified novel genes supporting infectious bronchitis virus (IBV) replication in human cells. The several host factors identified in this study may provide directions for future research on targeted therapeutics.
L Particles Transmit Viral Proteins from Herpes Simplex Virus 1-Infected Mature Dendritic Cells to Uninfected Bystander Cells, Inducing CD83 Downmodulation
Authors: Heilingloh Christiane S. , Kummer Mirko , Mühl-Zürbes Petra , Drassner Christina , Daniel Christoph , Klewer Monika , Steinkasserer Alexander
Published: August 2015

Mature dendritic cells (mDCs) are known as the most potent antigen-presenting cells (APCs) since they are also able to prime/induce naive T cells. Thus, mDCs play a pivotal role during the induction of antiviral immune responses. Remarkably, the cell surface molecule CD83, which was shown to have costimulatory properties, is targeted by herpes simplex virus 1 (HSV-1) for viral immune escape. Infection of mDCs with HSV-1 results in downmodulation of CD83, resulting in reduced T cell stimulation. In this study, we report that not only infected mDCs but also uninfected bystander cells in an infected culture show a significant CD83 reduction. We demonstrate that this effect is independent of phagocytosis and transmissible from infected to uninfected mDCs. The presence of specific viral proteins found in these uninfected bystander cells led to the hypothesis that viral proteins are transferred from infected to uninfected cells via L particles. These L particles are generated during lytic replication in parallel with full virions, called H particles. L particles contain viral proteins but lack the viral capsid and DNA. Therefore, these particles are not infectious but are able to transfer several viral proteins. Incubation of mDCs with L particles indeed reduced CD83 expression on uninfected bystander DCs, providing for the first time evidence that functional viral proteins are transmitted via L particles from infected mDCs to uninfected bystander cells, thereby inducing CD83 downmodulation.

IMPORTANCEHSV-1 has evolved a number of strategies to evade the host's immune system. Among others, HSV-1 infection of mDCs results in an inhibited T cell activation caused by degradation of CD83. Interestingly, CD83 is lost not only from HSV-1-infected mDCs but also from uninfected bystander cells. The release of so-called L particles, which contain several viral proteins but lack capsid and DNA, during infection is a common phenomenon observed among several viruses, such as human cytomegalovirus (HCMV), Epstein-Barr virus, and HSV-1. However, the detailed function of these particles is poorly understood. Here, we provide for the first time evidence that functional viral proteins can be transferred to uninfected bystander mDCs via L particles, revealing important biological functions of these particles during lytic replication. Therefore, the transfer of viral proteins by L particles to modulate uninfected bystander cells may represent an additional strategy for viral immune escape.
Oligomerization of Mumps Virus Phosphoprotein
Authors: Pickar Adrian , Elson Andrew , Yang Yang , Xu Pei , Luo Ming , He Biao
Published: August 2015

The mumps virus (MuV) genome encodes a phosphoprotein (P) that is important for viral RNA synthesis. P forms the viral RNA-dependent RNA polymerase with the large protein (L). P also interacts with the viral nucleoprotein (NP) and self-associates to form a homotetramer. The P protein consists of three domains, the N-terminal domain (P
N
), the oligomerization domain (P
O
), and the C-terminal domain (P
C
). While P
N
is known to relax the NP-bound RNA genome, the roles of P
O
and P
C
are not clear. In this study, we investigated the roles of P
O
and P
C
in viral RNA synthesis using mutational analysis and a minigenome system. We found that P
N
and P
C
functions can be
trans
-complemented. However, this complementation requires P
O
, indicating that P
O
is essential for P function. Using this
trans
-complementation system, we found that P forms parallel dimers (P
N
to P
N
and P
C
to P
C
). Furthermore, we found that residues R231, K238, K253, and K260 in P
O
are critical for P's functions. We identified P
C
to be the domain that interacts with L. These results provide structure-function insights into the role of MuV P.

IMPORTANCE
MuV, a paramyxovirus, is an important human pathogen. The P protein of MuV is critical for viral RNA synthesis. In this work, we established a novel minigenome system that allows the domains of P to be complemented in
trans
. Using this system, we confirmed that MuV P forms parallel dimers. An understanding of viral RNA synthesis will allow the design of better vaccines and the development of antivirals.
Characterization of the Interaction between the Matrix Protein of Vesicular Stomatitis Virus and the Immunoproteasome Subunit LMP2
Authors: Beilstein Frauke , Obiang Linda , Raux Hélène , Gaudin Yves
Published: August 2015

The matrix protein (M) of vesicular stomatitis virus (VSV) is involved in virus assembly, budding, gene regulation, and cellular pathogenesis. Using a yeast two-hybrid system, the M globular domain was shown to interact with LMP2, a catalytic subunit of the immunoproteasome (which replaces the standard proteasome catalytic subunit PSMB6). The interaction was validated by coimmunoprecipitation of M and LMP2 in VSV-infected cells. The sites of interaction were characterized. A single mutation of M (I96A) which significantly impairs the interaction between M and LMP2 was identified. We also show that M preferentially binds to the inactive precursor of LMP2 (bearing an N-terminal propeptide which is cleaved upon LMP2 maturation). Furthermore, taking advantage of a sequence alignment between LMP2 and its proteasome homolog, PSMB6 (which does not bind to M), we identified a mutation (L45R) in the S1 pocket where the protein substrate binds prior to cleavage and a second one (D17A) of a conserved residue essential for the catalytic activity, resulting in a reduction of the level of binding to M. The combination of both mutations abolishes the interaction. Taken together, our data indicate that M binds to LMP2 before its incorporation into the immunoproteasome. As the immunoproteasome promotes the generation of major histocompatibility complex (MHC) class I-compatible peptides, a feature which favors the recognition and the elimination of infected cells by CD8 T cells, we suggest that M, by interfering with the immunoproteasome assembly, has evolved a mechanism that allows infected cells to escape detection and elimination by the immune system.

IMPORTANCE
The immunoproteasome promotes the generation of MHC class I-compatible peptides, a feature which favors the recognition and the elimination of infected cells by CD8 T cells. Here, we report on the association of vesicular stomatitis virus (VSV) matrix protein (M) with LMP2, one of the immunoproteasome-specific catalytic subunits. M preferentially binds to the LMP2 inactive precursor. The M-binding site on LMP2 is facing inwards in the immunoproteasome and is therefore not accessible to M after its assembly. Hence, M binds to LMP2 before its incorporation into the immunoproteasome. We suggest that VSV M, by interfering with the immunoproteasome assembly, has evolved a mechanism that allows infected cells to escape detection and elimination by the immune system. Modulating this M-induced immunoproteasome impairment might be relevant in order to optimize VSV for oncolytic virotherapy.
A prediction model to estimate completeness of electronic physician claims databases
Authors: Lix Lisa M , Yao Xue , Kephart George , Quan Hude , Smith Mark , Kuwornu John Paul , Manoharan Nitharsana , Kouokam Wilfrid , Sikdar Khokan
Published: August 2015

Objectives

Electronic physician claims databases are widely used for chronic disease research and surveillance, but quality of the data may vary with a number of physician characteristics, including payment method. The objectives were to develop a prediction model for the number of prevalent diabetes cases in fee-for-service (FFS) electronic physician claims databases and apply it to estimate cases among non-FFS (NFFS) physicians, for whom claims data are often incomplete.

Design

A retrospective observational cohort design was adopted.

Setting

Data from the Canadian province of Newfoundland and Labrador were used to construct the prediction model and data from the province of Manitoba were used to externally validate the model.

Participants

A cohort of diagnosed diabetes cases was ascertained from physician claims, insured resident registry and hospitalisation records. A cohort of FFS physicians who were responsible for the diagnosis was ascertained from physician claims and registry data.

Primary and secondary outcome measures

A generalised linear model with a γ distribution was used to model the number of diabetes cases per FFS physician as a function of physician characteristics. The expected number of diabetes cases per NFFS physician was estimated.

Results

The diabetes case cohort consisted of 31 714 individuals; the mean cases per FFS physician was 75.5 (median=49.0). Sex and years since specialty licensure were significantly associated (p<0.05) with the number of cases per physician. Applying the prediction model to NFFS physician registry data resulted in an estimate of 18 546 cases; only 411 were observed in claims data. The model demonstrated face validity in an independent data set.

Conclusions

Comparing observed and predicted disease cases is a useful and generalisable approach to assess the quality of electronic databases for population-based research and surveillance.
Association of worker characteristics and early reimbursement for physical therapy, chiropractic and opioid prescriptions with workers’ compensation claim duration, for cases of acute low back pain: an observational cohort study
Authors: Busse Jason W , Ebrahim Shanil , Heels-Ansdell Diane , Wang Li , Couban Rachel , Walter Stephen D
Published: August 2015

Objective

To assess the association between early reimbursement for physiotherapy, chiropractic and opioid prescriptions for acute low back pain (LBP) with disability claim duration.

Design

Observational cohort study.

Setting and participants

From a random sample of 6665 claims for acute, uncomplicated LBP approved by the Ontario Workplace Safety and Insurance Board (WSIB) in 2005, we analysed 1442 who remained on full benefits at 4 weeks after claim approval.

Primary outcome measure

Our primary outcome was WSIB claim duration.

Results

We had complete data for all but 3 variables, which had <15% missing data, and we included missing data as a category for these factors. Our time-to-event analysis was adjusted for demographic, workplace and treatment factors, but not injury severity, although we attempted to include a sample with very similar, less-severe injuries. Regarding significant factors and treatment variables in our adjusted analysis, older age (eg, HR for age ≥55 vs <25=0.52; 99% CI 0.36 to 0.74) and WSIB reimbursement for opioid prescription in the first 4 weeks of a claim (HR=0.68; 99% CI 0.53 to 0.88) were associated with longer claim duration. Higher predisability income was associated with longer claim duration, but only among persistent claims (eg, HR for active claims at 1 year with a predisability income >$920 vs ≤$480/week=0.34; 99% CI 0.17 to 0.68). Missing data for union membership (HR=1.27; 99% CI 1.01 to 1.59), and working for an employer with a return-to-work programme were associated with fewer days on claim (HR=1.78; 99% CI 1.45 to 2.18). Neither reimbursement for physiotherapy (HR=1.01; 99% CI 0.86 to 1.19) nor chiropractic care (HR for active claims at 60 days=1.15; 99% CI 0.94 to 1.41) within the first 4 weeks was associated with claim duration. Our meta-analysis of 3 studies (n=51 069 workers) confirmed a strong association between early opioid use and prolonged claim duration (HR=0.57, 95% CI 0.48 to 0.69; low certainty evidence).

Conclusions

Our analysis found that early WSIB reimbursement for physiotherapy or chiropractic care, in claimants fully off work for more than 4 weeks, was not associated with claim duration, and that early reimbursement for opioids predicted prolonged claim duration. Well-designed randomised controlled trials are needed to verify our findings and establish causality between these variables and claim duration.
Treatment of experimental aneurysms with a new liquid embolic agent and a retrievable stent: proof of concept and feasibility study
Authors: Berenstein Alejandro
Published: August 2015

Background

Occlusion of canine bifurcation and sidewall aneurysms was undertaken with a new liquid embolic agent (PHIL 35) assisted by a high-density partially retrievable stent (FRED) with preservation of the carotid artery.

Methods

Three dogs were used as acute preparations for development of the technique and two were used for chronic studies lasting 90 days. In one animal we intentionally did not completely fill the aneurysm to determine the long-term results of incomplete treatment. The degree of occlusion, carotid artery compromise, and dislodgement and/or migration of embolic material in treated aneurysms were assessed.

Results

All aneurysms planned for complete obliteration were totally occluded successfully. By design, we partially occluded one aneurysm. In this aneurysm, angiography performed at 30 days revealed less filling, but at 90 days it had persistent small residual filling. We did not detect any distal embolization during the injection and no angiographic occlusions, change in configuration, or delayed migration of the embolic material were found. In the inspected stent, no foreign material was noted. In four animals we successfully removed the stent with preservation of the integrity of the carotid artery. In the fifth we intentionally left both stents deployed.

Conclusions

We have developed a new treatment for cerebral aneurysms using a combination of a retrievable stent and a new liquid embolic agent.