bluuebunny/crossref_metadata_2025 huggingface dataset:
A comparison of three infant skinfold reference standards: Tanner–Whitehouse, Cambridge Infant Growth Study, and WHO Child Growth Standards
Authors: Miller Elizabeth M.
Published: February 2014

As researchers increasingly focus on early infancy as a critical period of development, there is a greater need for methodological tools that can address all aspects of infant growth. Infant skinfold measures, in particular, are measurements in need of reliable reference standards that encompass all ages of infants and provide an accurate assessment of the relative fatness of a population. This report evaluates three published reference standards for infant skinfold measurements: Tanner–Whitehouse, Cambridge Infant Growth Study, and the World Health Organization (WHO) Child Growth Standards. To assess these standards, triceps skinfolds from a population of rural Kenyan infants (n = 250) and triceps skinfolds and subscapular skinfolds from infants in the National Health and Nutrition Examination Survey 1999–2002 (NHANES; n = 1197) were calculated as z‐scores from the lambda‐mu‐sigma curves provided by each reference population. The Tanner–Whitehouse standards represented both the Kenyan and US populations as lean, while the Cambridge standards represented both populations as overfat. The distribution of z‐scores based on the WHO standards fell in the middle, but excluded infants from both populations who were below the age of 3 months. Based on these results, the WHO reference standard is the best skinfold reference standard for infants over the age of 3 months. For populations with infants of all ages, the Tanner–Whitehouse standards are recommended, despite representing both study populations as underfat. Ideally, the WHO will extend their reference standard to include infants between the ages of 0 and 3 months.
Effectiveness of milk whey protein‐based ready‐to‐use therapeutic food in treatment of severe acute malnutrition in Malawian under‐5 children: a randomised, double‐blind, controlled non‐inferiority clinical trial
Authors: Bahwere Paluku , Banda Theresa , Sadler Kate , Nyirenda Gertrude , Owino Victor , Shaba Bina , Dibari Filippo , Collins Steve
Published: February 2014

The cost of ready‐to‐use therapeutic food (RUTF) used in community‐based management of acute malnutrition has been a major obstacle to the scale up of this important child survival strategy. The current standard recipe for RUTF [peanut‐based RUTF (P‐RUTF)] is made from peanut paste, milk powder, oil, sugar, and minerals and vitamins. Milk powder forms about 30% of the ingredients and may represent over half the cost of the final product. The quality of whey protein concentrates 34% (WPC34) is similar to that of dried skimmed milk (DSM) used in the standard recipe and can be 25–33% cheaper. This blinded, parallel group, randomised, controlled non‐inferiority clinical trial tested the effectiveness in treating severe acute malnutrition (SAM) of a new RUTF formulation WPC‐RUTF in which WPC34 was used to replace DSM. Average weight gain (non‐inferiority margin Δ = −1.2 g kg−1 day−1) and recovery rate (Δ = −10%) were the primary outcomes, and length of stay (LOS) was the secondary outcome (Δ = +14 days). Both per‐protocol (PP) and intention‐to‐treat (ITT) analyses showed that WPC‐RUTF was not inferior to P‐RUTF for recovery rate [difference and its 95% confidence interval (CI) of 0.5% (95% CI –2.7, 3.7) in PP analysis and 0.6% (95% CI –5.2, 6.3) in ITT analysis] for average weight gain [0.2 (−0.5; 0.9) for both analyses] and LOS [−1.6 days (95% CI, −4.6, 1.4 days) in PP analysis and −1.9 days (95% CI, −4.6, 0.8 days) for ITT analysis]. In conclusion, whey protein‐based RUTF is an effective cheaper alternative to the standard milk‐based RUTF for the treatment of SAM.
A genetic approach to study H2O2 scavenging in fission yeast – distinct roles of peroxiredoxin and catalase
Authors: Paulo Esther , García‐Santamarina Sarela , Calvo Isabel A. , Carmona Mercè , Boronat Susanna , Domènech Alba , Ayté José , Hidalgo Elena
Published: February 2014

The main peroxiredoxin in Schizosaccharomyces pombe, Tpx1, is important to sustain aerobic growth, and cells lacking this protein are only able to grow on solid plates under anaerobic conditions. We have found that deletion of the gene coding for thioredoxin reductase, trr1, is a suppressor of the sensitivity to aerobic growth of Δtpx1 cells, so that cells lacking both proteins are able to grow on solid plates in the presence of oxygen. We have investigated this suppression effect, and determined that it depends on the presence of catalase, which is constitutively expressed in Δtrr1 cells in a transcription factor Pap1‐dependent manner. A complete characterization of the repertoire of hydrogen peroxide scavenging activities in fission yeast suggests that Tpx1 is the only enzyme with sufficient sensitivity for peroxides and cellular abundance as to control the low levels produced during aerobic growth, catalase being the next barrier of detoxification when the steady‐state levels of peroxides are increased in Δtpx1 cells. Gpx1, the only glutathione peroxidase encoded by the S. pombe genome, only has a minor secondary role when extracellular peroxides are added. Our study proposes non‐overlapping roles for the different hydrogen peroxide scavenging activities of this eukaryotic organism.
Sequence diversity in the large subunit of RNA polymerase I contributes to Mefenoxam insensitivity in Phytophthora infestans
Authors: Randall Eva , Young Vanessa , Sierotzki Helge , Scalliet Gabriel , Birch Paul R. J. , Cooke David E. L. , Csukai Michael , Whisson Stephen C.
Published: February 2014

Phenylamide fungicides have been widely used for the control of oomycete‐incited plant diseases for over 30 years. Insensitivity to this chemical class of fungicide was recorded early in its usage history, but the precise protein(s) conditioning insensitivity has proven difficult to determine. To determine the genetic basis of insensitivity and to inform strategies for the cloning of the gene(s) responsible, genetic crosses were established between Mefenoxam sensitive and intermediate insensitive isolates of Phytophthora infestans, the potato late blight pathogen. F1 progeny showed the expected semi‐dominant phenotypes for Mefenoxam insensitivity and suggested the involvement of multiple loci, complicating the positional cloning of the gene(s) conditioning insensitivity to Mefenoxam. Instead, a candidate gene strategy was used, based on previous observations that the primary effect of phenylamide compounds is to inhibit ribosomal RNA synthesis. The subunits of RNA polymerase I (RNApolI) were sequenced from sensitive and insensitive isolates and F1 progeny. Single nucleotide polymorphisms (SNPs) specific to insensitive field isolates were identified in the gene encoding the large subunit of RNApolI. In a survey of field isolates, SNP T1145A (Y382F) showed an 86% association with Mefenoxam insensitivity. Isolates not showing this association belonged predominantly to one P. infestans genotype. The transfer of the ‘insensitive’ allele of RPA190 to a sensitive isolate yielded transgenic lines that were insensitive to Mefenoxam. These results demonstrate that sequence variation in RPA190 contributes to insensitivity to Mefenoxam in P. infestans.
Motility response to colonic distention is increased in postinfectious irritable bowel syndrome (PI‐IBS)
Authors: Kanazawa M. , Palsson O. S. , van Tilburg M. A. L. , Gangarosa L. M. , Fukudo S. , Whitehead W. E.
Published: February 2014

Background

Acute intestinal infection leads to persistent intestinal smooth muscle hypercontractility and pain hypersensitivity after resolution of the infection in animal models. We investigated whether postinfectious irritable bowel syndrome (PI‐IBS) is associated with abnormalities in phasic contractions of the colon, smooth muscle tone, and pain sensitivity compared to non‐PI‐IBS (NI‐IBS) or healthy controls (HC).

Methods

Two hundred and eighteen Rome III‐positive IBS patients and 43 HC participated. IBS patients were designated PI‐IBS, if their IBS symptoms began following an episode of gastroenteritis characterized by two or more of: fever, vomiting, or diarrhea. Pain threshold to phasic distentions of the descending colon was assessed using a barostat. Colonic motility was assessed with the barostat bag minimally inflated to the individual operating pressure (IOP), at 20 mmHg above the IOP, and following a test meal. IBS symptom severity and psychological symptoms were assessed by the IBS Severity Scale (IBS‐SS) and the Brief Symptom Inventory‐18 (BSI‐18).

Key Results

Twenty two (10.1%) met criteria for PI‐IBS. Both IBS and HC groups showed a significant increase in motility index during intraluminal distention and following meals. The magnitude of the response to distention above (orad to) the balloon was significantly greater in PI‐IBS compared with NI‐IBS (p < 0.05) or HC (p < 0.01). Differences between PI‐IBS and NI‐IBS were not significant for IBS symptom severity, pain threshold, barostat bag volumes, or any psychological score on the BSI‐18.

Conclusions & Inferences

Patients with PI‐IBS have greater colonic hypercontractility than NI‐IBS. We speculate that sustained mild mucosal inflammation may cause this colonic irritability.
Impairment on theory of mind and empathy in patients with stroke
Authors: Yeh Zai‐Ting , Tsai Chung‐Fen
Published: February 2014

Aims

Impaired social function has been described in patients following stroke. The present study was designed to explore the degree of impairment in the ability to infer mental states in others, or cognitive and affective theory of mind, and empathy, in patients with stroke.

Method

A total of 34 patients with stroke were compared to 40 control subjects on tasks testing verbal and non‐verbal theory of mind and empathy.

Results

Results indicated that patients with stroke were significantly impaired in both cognitive and affective theory of mind, even controlling for basic cognitive function and emotional processing. The patients with right stroke had poorer performance than those with left stroke on the cognitive component of non‐verbal theory of mind. On the subscale of cognitive empathy, the right stroke group had poorer performance on perspective‐taking than the control group.

Conclusion

The right hemisphere may play an important role in decoding non‐verbal cues to infer others' minds as well as the processing of empathy, especially the ability of perspective‐taking.
Reducing the stigma of depression through neurobiology‐based psychoeducation: A randomized controlled trial
Authors: Han Der‐Yan , Chen Sue‐Huei
Published: February 2014

Aims

Attribution theory claims that people who are stigmatized experience more negative emotional and behavioral reactions from others when they are thought to be responsible for their problems. Accordingly, this study proposed a neurobiology‐based psychoeducational intervention, which attempted to reduce people's blameworthy attitudes toward and social distance from depressed individuals.

Methods

One hundred and thirty‐two college students were randomly assigned to an experimental and control group. Participants in the experimental group received a 30‐min lecture on neurobiology‐based psychoeducation for depressive disorders, and were asked to fill out questionnaires before and 2 weeks after the intervention. The control group, with no intervention, also filled out the same questionnaires before and 2 weeks after the experiment. The main contents of the neurobiology‐based psychoeducation concerned the neurotransmission processes and biological mechanisms of depression, in order to emphasize the biological attribution of depression.

Results

An ancova indicated that the neurobiology‐based psychoeducational intervention significantly elevated the biological attribution of depression and reduced the social distance from depressed individuals. Psychological blameworthy attitudes toward depression, however, did not significantly change.

Conclusions

Through a brief psychoeducation program about depression, knowledge of neuroscience could lead to positive benefits. Public awareness that depression can be effectively prevented and treated may be a way in which people can accept depressed individuals. Further studies are needed to certify the mechanisms of the effect of neurobiology‐based psychoeducation.
REER Imbalances and Macroeconomic Adjustments in the Proposed West African Monetary Union
Authors: Asongu Simplice
Published: February 2014

With the spectre of the euro crisis hunting embryonic monetary unions, we use a dynamic model of a small open economy to analyse real effective exchange rate (REER) imbalances and examine whether the movements in the aggregate real exchange rates are consistent with the underlying macroeconomic fundamentals in the proposed West African Monetary Union (WAMU). Using both country‐oriented and WAMU panel‐based specifications, we show that the long‐run behaviour of the REERs can be explained by fluctuations in the terms of trade, productivity, investment, debt and openness. While there is still significant evidence of cross‐country differences in the relationship between underlying macroeconomic fundamentals and corresponding REERs, the embryonic WAMU has a stable error correction mechanism, with four of the five cointegration relations having signs that are consistent with the predictions from economic theory. Policy implications are discussed, and the conclusions of the analysis are a valuable contribution to the scholarly and policy debate over whether the creation of a sustainable monetary union should precede convergence in macroeconomic fundamentals that determine REER adjustments.
Monty's dilemma with no formulas
Authors: Falk Ruma
Published: February 2014

The article presents an attempt to analyse Monty's dilemma by means of conversational formula‐free dialogues and to simulate the problem by composing isomorphic stories. The crucial roles of specifying the underlying scenarios and explicating epistemic and probabilistic assumptions are highlighted.
The New Business of Football: A Study of Current and Aspirant Football Club Managers
Authors: Morrow Stephen , Howieson Brian
Published: February 2014

Professional football (soccer) in Europe has changed dramatically in the past two decades, largely due to the escalation of media rights deals. Many professional football clubs are now complex businesses, intrinsically concerned with financial matters. Within the rapidly changing business context of football, the aim of this research is to further understand the main issues that are related to a career as a manager. This paper has five sections: (1) we offer an appraisal of the general literature as it applies to professional football management; (2) we introduce the theoretical focus of the article with specific reference to the “career” and describe the context and background to the research; (3) we describe the research methodology and present and discuss the research results, which center on the career development of the manager; the position of a manager in organizational structures, and how the changing organization affects the role of manager; (4) we set out the conclusions and implications of our research; and (5) we offer our plans to progress this research, enabling a new body of knowledge to be developed on this specialized role.
Nanoporous BiVO
4
Photoanodes with Dual-Layer Oxygen Evolution Catalysts for Solar Water Splitting
Authors: Kim Tae Woo , Choi Kyoung-Shin
Published: February 2014

A Boost for Bismuth Vanadate

In theory, given its light-absorption spectrum, bismuth vanadate should be an effective photoanode for solar water-splitting. However, in prior studies, few of the “holes” generated upon photoexcitation have persisted long enough to strip electrons from water.
Kim and Choi
(p.
990
, published online 13 February) now show that the use of a hydrophobic vanadium source in the semiconductor's synthesis results in a high-surface-area morphology with substantially enhanced hole lifetimes. Deposition of two successive catalyst layers enhanced the proportion of holes that reacted with water at the surface, thereby raising the efficiency of the oxygen evolution reaction.
Trained Immunity or Tolerance: Opposing Functional Programs Induced in Human Monocytes after Engagement of Various Pattern Recognition Receptors
Authors: Ifrim Daniela C. , Quintin Jessica , Joosten Leo A. B. , Jacobs Cor , Jansen Trees , Jacobs Liesbeth , Gow Neil A. R. , Williams David L. , van der Meer Jos W. M. , Netea Mihai G.
Published: February 2014

Upon priming withCandida albicansor with the fungal cell wall component β-glucan, monocytes respond with an increased cytokine production upon restimulation, a phenomenon termed “trained immunity.” In contrast, the prestimulation of monocytes with lipopolysaccharide has long been known to induce tolerance. Because the vast majority of commensal microorganisms belong to bacterial or viral phyla, we sought to systematically investigate the functional reprogramming of monocytes induced by the stimulation of pattern recognition receptors (PRRs) with various bacterial or viral ligands. Monocytes were functionally programmed for either enhanced (training) or decreased (tolerance) cytokine production, depending on the type and concentration of ligand they encountered. The functional reprogramming of monocytes was also associated with cell shape, granulocity, and cell surface marker modifications. The training effect required p38- and Jun N-terminal protein kinase (JNK)-mediated mitogen-activated protein kinase (MAPK) signaling, with specific signaling patterns directing the functional fate of the cell. The long-term effects on the function of monocytes were mediated by epigenetic events, with both histone methylation and acetylation inhibitors blocking the training effects. In conclusion, our experiments identify the ability of monocytes to acquire adaptive characteristics after prior activation with a wide variety of ligands. Trained immunity and tolerance are two distinct and opposing functional programs induced by the specific microbial ligands engaging the monocytes.
Evaluation of Gamma Interferon Immune Response Elicited by the Newly Constructed PstS-1(285-374):CFP10 Fusion Protein To Detect Mycobacterium tuberculosis Infection
Authors: Araujo Leonardo Silva de , Mello Fernanda Carvalho de Queiroz , da Silva Nidai de Bárbara Moreira , Leung Janaina Aparecida Medeiros , Machado Silvia Maria Almeida , Sardella Isabela Gama , Maciel Renata de Moraes , Saad Maria Helena Féres
Published: February 2014

The PstS1 antigen is highly immunogenic, principally when combined with CFP10 during both latent and active TB infection. In the present study, a selected
pstS1
gene fragment was cloned, fused with CFP10, and expressed in
Escherichia coli
. The product [PstS-1(285-374):CFP10] was compared to the recombinant fused RD1 (region of deletion 1) protein (ESAT-6:CFP10) in detecting
Mycobacterium tuberculosis
infection in 108 recent contacts of pulmonary tuberculosis (TB) cases, considering a positive tuberculin skin test (TST) to be the baseline. The release of gamma interferon (IFN-γ) in 22-h whole-blood and 5-day lymphocyte stimulation assays primed with each antigen was determined. All contacts were clinically followed for up to 1 year, and 87% of the tuberculin skin test-positive (TST
positive
) patients accepted preventative treatment. Concerning the IFN-γ response to PstS-1(285-374):CFP10 in the 22-h and 5-day assays, a slight increase in contact-TST
positive
detection was observed (23/54 and 26/54) compared to the level seen with the RD1 protein (18/54 and 24/54) whereas in the TST
negative
group, similarly lower numbers (≤5/48) of responders were achieved for both antigens, except for RD1 in the 5-day assay (8/48). By combining the IFN-γ responders to both antigens in the 5-day assays, slightly higher increases in positivity were found in the TST
positive
(32/54) and TST
negative
(10/48) groups. Two of 12 untreated TST
positive
contacts progressed to active TB and were concordantly positive in all assays, except for one contact who lacked positivity in the RD1 5-day assay. We demonstrated for the first time that PstS-1(285-374):CFP10 slightly increased contact positivity and detection of active disease progression, suggesting its potential application as a TB infection marker.
Molecular Epidemiology of Merkel Cell Polyomavirus: Evidence for Geographically Related Variant Genotypes
Authors: Martel-Jantin Claire , Filippone Claudia , Tortevoye Patricia , Afonso Philippe V. , Betsem Edouard , Descorps-Declere Stéphane , Nicol Jérôme T. J. , Touzé Antoine , Coursaget Pierre , Crouzat Maryse , Berthet Nicolas , Cassar Olivier , Gessain Antoine
Published: February 2014

Merkel cell polyomavirus (MCPyV) is linked to a cutaneous cancer mainly occurring in Caucasians. DNA from skin swabs of 255 adults, originating from the 5 continents, were subjected to MCPyV PCRs. Phylogenetic analyses demonstrate the existence of 5 major geographically related MCPyV genotypes (Europe/North America, Africa [sub-Saharan], Oceania, South America, and Asia/Japan).
Multilocus Microsatellite Genotyping Array for Investigation of Genetic Epidemiology of Pneumocystis jirovecii
Authors: Parobek Christian M. , Jiang Linda Y. , Patel Jaymin C. , Alvarez-Martínez Miriam J. , Miro Jose M. , Worodria William , Andama Alfred , Fong Serena , Huang Laurence , Meshnick Steven R. , Taylor Steve M. , Juliano Jonathan J.
Published: February 2014

Pneumocystis jirovecii
is a symbiotic respiratory fungus that causes pneumonia (PcP) in immunosuppressed patients. Because
P. jirovecii
cannot be reliably cultured
in vitro
, it has proven difficult to study and gaps in our understanding of the organism persist. The release of a draft genome for the organism opens the door for the development of new genotyping approaches for studying its molecular epidemiology and global population structure. We identified and validated 8 putatively neutral microsatellite markers and 1 microsatellite marker linked to the dihydropteroate synthase gene (
dhps
), the enzymatic target of sulfa drugs used for PcP prevention and treatment. Using these tools, we analyzed
P. jirovecii
isolates from HIV-infected patients from three geographically distant populations: Uganda, the United States, and Spain. Among the 8 neutral markers, we observed high levels of allelic heterozygosity (average H
e
, 0.586 to 0.842). Consistent with past reports, we observed limited global population structuring, with only the Ugandan isolates showing minor differentiation from the other two populations. In Ugandan isolates that harbored mutations in
dhps
, the microsatellite locus linked to
dhps
demonstrated a depressed H
e
, consistent with positive directional selection for sulfa resistance mutations. Using a subset of these microsatellites, analyses of individual and paired samples from infections in San Francisco, CA, showed reliable typeability within a single infection and high discriminatory power between infections. These features suggest that this novel microsatellite typing approach will be an effective tool for molecular-epidemiological investigations into
P. jirovecii
population structure, transmission, and drug resistance.
Disseminated Infection with Mycobacterium tilburgii in a Male Immunocompromised Patient
Authors: Temmerman Sarah , Vandekerckhove Linos , Sermijn Erica , Vogelaers Dirk , Claeys Geert , Vaneechoutte Mario , Cools Piet , Callens Steven
Published: February 2014

Mycobacterium tilburgii
is a nonculturable nontuberculous mycobacterium identifiable only by molecular methods. We report a case of disseminated
M. tilburgii
infection illustrating the importance of 16S rRNA gene sequencing to determine the responsible mycobacterial pathogen and the difficulties in tailoring antimycobacterial treatment in the absence of a culturable organism.
Real-Time PCR Assay for Rapid Detection of Epidemiologically and Clinically Significant Mycobacterium tuberculosis Beijing Genotype Isolates
Authors: Mokrousov Igor , Vyazovaya Anna , Zhuravlev Viacheslav , Otten Tatiana , Millet Julie , Jiao Wei-Wei , Shen A-Dong , Rastogi Nalin , Vishnevsky Boris , Narvskaya Olga
Published: February 2014

Mycobacterium tuberculosis
Beijing genotype strains are rapidly disseminating, frequently hypervirulent, and multidrug resistant. Here, we describe a method for their rapid detection by real-time PCR that targets the specific IS
6110
insertion in the
dnaA-dnaN
genome region. The method was evaluated with a geographically and genetically diverse collection representing areas in East Asia and the former Soviet Union in which the Beijing genotype is endemic and epidemic (i.e., major foci of its global propagation) and with clinical specimens.
Concurrent Infection of the Urinary Tract with Encephalitozoon cuniculi and Enterocytozoon bieneusi in a Renal Transplant Recipient
Authors: Kicia Marta , Wesolowska Maria , Jakuszko Katarzyna , Kopacz Zaneta , Sak Bohumil , Květonová Dana , Krajewska Magdalena , Kváč Martin
Published: February 2014

A urinary tract coinfection, caused by
Encephalitozoon cuniculi
genotype II and
Enterocytozoon bieneusi
genotype D, was identified in an HIV-seronegative renal transplant recipient kept under lifelong immunosuppression. To our knowledge, this is the first report describing concurrent infection with these two microsporidia species in organ transplant recipients.
Serological and Molecular Diagnosis of Hepatitis Delta Virus Infection: Results of a French National Quality Control Study
Authors: Brichler Ségolène , Le Gal Frédéric , Neri-Pinto Fernando , Mansour Wael , Roulot Dominique , Laperche Syria , Gordien Emmanuel
Published: February 2014

A French national quality control study for the serological and molecular diagnosis of hepatitis delta virus (HDV) was organized. Total HDV antibodies were properly detected by all laboratories; 8/14 laboratories failed to detect low titers of IgM, and 6/11 failed to quantify and/or underestimated the RNA viral load in several samples. These discrepancies are likely related to the molecular diversity of HDV.
Identifying Neisseria Species by Use of the 50S Ribosomal Protein L6 (
rplF
) Gene
Authors: Bennett Julia S. , Watkins Eleanor R. , Jolley Keith A. , Harrison Odile B. , Maiden Martin C. J.
Published: February 2014

The comparison of 16S rRNA gene sequences is widely used to differentiate bacteria; however, this gene can lack resolution among closely related but distinct members of the same genus. This is a problem in clinical situations in those genera, such as
Neisseria
, where some species are associated with disease while others are not. Here, we identified and validated an alternative genetic target common to all
Neisseria
species which can be readily sequenced to provide an assay that rapidly and accurately discriminates among members of the genus. Ribosomal multilocus sequence typing (rMLST) using ribosomal protein genes has been shown to unambiguously identify these bacteria. The PubMLST
Neisseria
database (
http://pubmlst.org/neisseria/
) was queried to extract the 53 ribosomal protein gene sequences from 44 genomes from diverse species. Phylogenies reconstructed from these genes were examined, and a single 413-bp fragment of the 50S ribosomal protein L6 (
rplF
) gene was identified which produced a phylogeny that was congruent with the phylogeny reconstructed from concatenated ribosomal protein genes. Primers that enabled the amplification and direct sequencing of the
rplF
gene fragment were designed to validate the assay
in vitro
and
in silico
. Allele sequences were defined for the gene fragment, associated with particular species names, and stored on the PubMLST
Neisseria
database, providing a curated electronic resource. This approach provides an alternative to 16S rRNA gene sequencing, which can be readily replicated for other organisms for which more resolution is required, and it has potential applications in high-resolution metagenomic studies.
Outbreak of Vancomycin-Susceptible Enterococcus faecium Containing the Wild-Type
vanA
Gene
Authors: Szakacs Tom A. , Kalan Lindsay , McConnell Michael J. , Eshaghi Alireza , Shahinas Dea , McGeer Allison , Wright Gerry D. , Low Donald E. , Patel Samir N.
Published: February 2014

Accurate detection of vancomycin-resistant enterococci (VRE) is essential in preventing transmission in health care settings. Chromogenic media are widely used for screening VRE because of fast turnaround times (TAT) and high sensitivity. We report an outbreak of
Enterococcus faecium
bearing
vanA
yet susceptible to vancomycin (vancomycin-variable
Enterococcus
[VVE]). Between October 2009 to March 2011, clinical and screening specimens (
n
= 14,747) were screened for VRE using VRE-selective medium and/or PCR. VVE isolates were genotyped to determine relatedness. Plasmids from these isolates were characterized by sequencing. Overall, 52 VVE isolates were identified, comprising 15% of all VRE isolates identified. Isolates demonstrated growth on Brilliance VRE agar (Oxoid) at 24 h of incubation but did not grow on brain heart infusion agar with 6 μg/ml vancomycin (Oxoid) or bile esculin azide agar with 6 μg/ml vancomycin (Oxoid) and were susceptible to vancomycin. Genotyping of 20 randomly selected VVE isolates revealed that 15/20 were identical, while 5 were highly related. PCR of the VVE transposon confirmed the presence of
vanHAXY
gene cluster; however,
vanS
(sensor) and
vanR
(regulator) genes were absent. The outbreak was controlled through routine infection control measures. We report an emergence of a fit strain of
E. faecium
containing
vanA
yet susceptible to vancomycin. Whether this new strain represents VRE has yet to be determined; however, unique testing procedures are required for reliable identification of VVE.
Herpes Simplex Virus 1 UL47 Interacts with Viral Nuclear Egress Factors UL31, UL34, and Us3 and Regulates Viral Nuclear Egress
Authors: Liu Zhuoming , Kato Akihisa , Shindo Keiko , Noda Takeshi , Sagara Hiroshi , Kawaoka Yoshihiro , Arii Jun , Kawaguchi Yasushi
Published: February 2014

Herpesviruses have evolved a unique mechanism for nuclear egress of nascent progeny nucleocapsids: the nucleocapsids bud through the inner nuclear membrane into the perinuclear space between the inner and outer nuclear membranes (primary envelopment), and enveloped nucleocapsids then fuse with the outer nuclear membrane to release nucleocapsids into the cytoplasm (de-envelopment). We have shown that the herpes simplex virus 1 (HSV-1) major virion structural protein UL47 (or VP13/VP14) is a novel regulator for HSV-1 nuclear egress. In particular, we demonstrated the following: (i) UL47 formed a complex(es) with HSV-1 proteins UL34, UL31, and/or Us3, which have all been reported to be critical for viral nuclear egress, and these viral proteins colocalized at the nuclear membrane in HSV-1-infected cells; (ii) the UL47-null mutation considerably reduced primary enveloped virions in the perinuclear space although capsids accumulated in the nucleus; and (iii) UL47 was detected in primary enveloped virions in the perinuclear space by immunoelectron microscopy. These results suggested that UL47 promoted HSV-1 primary envelopment, probably by interacting with the critical HSV-1 regulators for viral nuclear egress and by modulating their functions.

IMPORTANCE
Like other herpesviruses, herpes simplex virus 1 (HSV-1) has evolved a vesicle-mediated nucleocytoplasmic transport mechanism for nuclear egress of nascent progeny nucleocapsids. Although previous reports identified and characterized several HSV-1 and cellular proteins involved in viral nuclear egress, complete details of HSV-1 nuclear egress remain to be elucidated. In this study, we have presented data suggesting (i) that the major HSV-1 virion structural protein UL47 (or VP13/VP14) formed a complex with known viral regulatory proteins critical for viral nuclear egress and (ii) that UL47 played a regulatory role in HSV-1 primary envelopment. Thus, we identified UL47 as a novel regulator for HSV-1 nuclear egress.
17β-Estradiol Protects Females against Influenza by Recruiting Neutrophils and Increasing Virus-Specific CD8 T Cell Responses in the Lungs
Authors: Robinson Dionne P. , Hall Olivia J. , Nilles Tricia L. , Bream Jay H. , Klein Sabra L.
Published: February 2014

17β-Estradiol (E2) treatment limits the pathology associated with pulmonary diseases caused by pathogens, allergens, and asthma, partly by reducing the production of proinflammatory cytokines and chemokines. To test the hypothesis that E2 protects against influenza A virus (IAV) infection by altering the recruitment and activity of innate immune cells and T cells, chemokine concentrations were measured and innate and adaptive immune cells were enumerated from the lungs of E2- and placebo-treated ovariectomized female C57BL/6 mice following infection. Females treated with E2 experienced less morbidity but had similar lung virus titers to placebo-treated females. Females treated with E2 had lower induction of CCL2 but higher CCL3 and CXCL1 responses in their lungs than placebo-treated females. Pulmonary recruitment of neutrophils, NK cells, macrophages, and dendritic cells was increased following infection, but only neutrophil numbers were greater in E2-treated than placebo-treated females. Neutrophils enhance the responses of influenza virus-specific CD8 T cells to promote virus clearance and improve the outcome of infection. Total numbers of virus-specific CD8 T cells were not altered by treatment with E2, but the proportion of gamma interferon (IFN-γ)- and tumor necrosis factor alpha (TNF-α)-producing, virus-specific CD8 T cells was increased. Neutrophil depletion in E2-treated females increased morbidity, reduced pulmonary production of chemoattractants for neutrophils, and reduced IFN-γ production by virus-specific CD8 T cells. Neutrophils mediate both inflammation and tissue repair during IAV infection and are regulated by E2 to improve the outcome of influenza in females.

IMPORTANCESevere influenza is associated with excessive inflammation that leads to tissue damage. We demonstrate that estradiol (E2) is a potent anti-inflammatory hormone that reduces the severity of influenza A virus infection in females. Treatment of female C57BL/6 mice with E2 does not affect virus replication but rather alters the production of chemokines, pulmonary recruitment of neutrophils, and the cytokine responses of virus-specific CD8 T cells to protect females against severe influenza.
Structural Stability of Influenza A(H1N1)pdm09 Virus Hemagglutinins
Authors: Yang Hua , Chang Jessie C. , Guo Zhu , Carney Paul J. , Shore David A. , Donis Ruben O. , Cox Nancy J. , Villanueva Julie M. , Klimov Alexander I. , Stevens James
Published: February 2014

The noncovalent interactions that mediate trimerization of the influenza hemagglutinin (HA) are important determinants of its biological activities. Recent studies have demonstrated that mutations in the HA trimer interface affect the thermal and pH sensitivities of HA, suggesting a possible impact on vaccine stability (). We used size exclusion chromatography analysis of recombinant HA ectodomain to compare the differences among recombinant trimeric HA proteins from early 2009 pandemic H1N1 viruses, which dissociate to monomers, with those of more recent virus HAs that can be expressed as trimers. We analyzed differences among the HA sequences and identified intermolecular interactions mediated by the residue at position 374 (HA0 numbering) of the HA2 subdomain as critical for HA trimer stability. Crystallographic analyses of HA from the recent H1N1 virus A/Washington/5/2011 highlight the structural basis for this observed phenotype. It remains to be seen whether more recent viruses with this mutation will yield more stable vaccines in the future.

IMPORTANCE
Hemagglutinins from the early 2009 H1N1 pandemic viruses are unable to maintain a trimeric complex when expressed in a recombinant system. However, HAs from 2010 and 2011 strains are more stable, and our work highlights that the improvement in stability can be attributed to an E374K substitution in the HA2 subunit of the stalk that emerged naturally in the circulating viruses.
Enhancement of Blood-Brain Barrier Permeability and Reduction of Tight Junction Protein Expression Are Modulated by Chemokines/Cytokines Induced by Rabies Virus Infection
Authors: Chai Qingqing , He Wen Q. , Zhou Ming , Lu Huijun , Fu Zhen F.
Published: February 2014

Infection with laboratory-attenuated rabies virus (RABV) enhances blood-brain barrier (BBB) permeability, which has been demonstrated to be an important factor for host survival, since it allows immune effectors to enter the central nervous system (CNS) and clear RABV. To probe the mechanism by which RABV infection enhances BBB permeability, the expression of tight junction (TJ) proteins in the CNS was investigated following intracranial inoculation with laboratory-attenuated or wild-type (wt) RABV. BBB permeability was significantly enhanced in mice infected with laboratory-attenuated, but not wt, RABV. The expression levels of TJ proteins (claudin-5, occludin, and zonula occludens-1) were decreased in mice infected with laboratory-attenuated, but not wt, RABV, suggesting that enhancement of BBB permeability is associated with the reduction of TJ protein expression in RABV infection. RABV neither infects the brain microvascular endothelial cells (BMECs) nor modulates the expression of TJ proteins in BMECs. However, brain extracts prepared from mice infected with laboratory-attenuated, but not wt, RABV reduced TJ protein expression in BMECs. It was found that brain extracts from mice infected with laboratory-attenuated RABV contained significantly higher levels of inflammatory chemokines/cytokines than those from mice infected with wt RABV. Pathway analysis indicates that gamma interferon (IFN-γ) is located in the center of the cytokine network in the RABV-infected mouse brain, and neutralization of IFN-γ reduced both the disruption of BBB permeabilityin vivoand the downregulation of TJ protein expressionin vitro. These findings indicate that the enhancement of BBB permeability and the reduction of TJ protein expression are due not to RABV infectionper sebut to virus-induced inflammatory chemokines/cytokines.

IMPORTANCEPrevious studies have shown that infection with only laboratory-attenuated, not wild-type, rabies virus (RABV) enhances blood-brain barrier (BBB) permeability, allowing immune effectors to enter the central nervous system (CNS) and clear RABV from the CNS. This study investigated the mechanism by which RABV infection enhances BBB permeability. It was found that RABV infection enhances BBB permeability by downregulation of tight junction (TJ) protein expression in the brain microvasculature. It was further found that it is not RABV infectionper sebut the chemokines/cytokines induced by RABV infection that downregulate the expression of TJ proteins and enhance BBB permeability. Blocking some of these cytokines, such as IFN-γ, ameliorated both the disruption of BBB permeability and the downregulation of TJ protein expression. These studies may provide a foundation for developing therapeutics for clinical rabies, such as medication that could be used to enhance BBB permeability.
Limited Hepatitis B Virus Replication Space in the Chronically Hepatitis C Virus-Infected Liver
Authors: Wieland S. F. , Asabe S. , Engle R. E. , Purcell R. H. , Chisari F. V.
Published: February 2014

We compared the kinetics and magnitude of hepatitis B virus (HBV) infection in hepatitis C virus (HCV)-naive and chronically HCV-infected chimpanzees in whose livers type I interferon-stimulated gene (ISG) expression is strongly induced. HBV infection was delayed and attenuated in the HCV-infected animals, and the number of HBV-infected hepatocytes was drastically reduced. These results suggest that establishment of HBV infection and its replication space is limited by the antiviral effects of type I interferon in the chronically HCV-infected liver.
Middle East Respiratory Syndrome Coronavirus 4a Protein Is a Double-Stranded RNA-Binding Protein That Suppresses PACT-Induced Activation of RIG-I and MDA5 in the Innate Antiviral Response
Authors: Siu Kam-Leung , Yeung Man Lung , Kok Kin-Hang , Yuen Kit-San , Kew Chun , Lui Pak-Yin , Chan Chi-Ping , Tse Herman , Woo Patrick C. Y. , Yuen Kwok-Yung , Jin Dong-Yan
Published: February 2014

Middle East respiratory syndrome coronavirus (MERS-CoV) is an emerging pathogen that causes severe disease in human. MERS-CoV is closely related to bat coronaviruses HKU4 and HKU5. Evasion of the innate antiviral response might contribute significantly to MERS-CoV pathogenesis, but the mechanism is poorly understood. In this study, we characterized MERS-CoV 4a protein as a novel immunosuppressive factor that antagonizes type I interferon production. MERS-CoV 4a protein contains a double-stranded RNA-binding domain capable of interacting with poly(I·C). Expression of MERS-CoV 4a protein suppressed the interferon production induced by poly(I·C) or Sendai virus. RNA binding of MERS-CoV 4a protein was required for IFN antagonism, a property shared by 4a protein of bat coronavirus HKU5 but not by the counterpart in bat coronavirus HKU4. MERS-CoV 4a protein interacted with PACT in an RNA-dependent manner but not with RIG-I or MDA5. It inhibited PACT-induced activation of RIG-I and MDA5 but did not affect the activity of downstream effectors such as RIG-I, MDA5, MAVS, TBK1, and IRF3. Taken together, our findings suggest a new mechanism through which MERS-CoV employs a viral double-stranded RNA-binding protein to circumvent the innate antiviral response by perturbing the function of cellular double-stranded RNA-binding protein PACT. PACT targeting might be a common strategy used by different viruses, including Ebola virus and herpes simplex virus 1, to counteract innate immunity.

IMPORTANCE
Middle East respiratory syndrome coronavirus (MERS-CoV) is an emerging and highly lethal human pathogen. Why MERS-CoV causes severe disease in human is unclear, and one possibility is that MERS-CoV is particularly efficient in counteracting host immunity, including the sensing of virus invasion. It will therefore be critical to clarify how MERS-CoV cripples the host proteins that sense viruses and to compare MERS-CoV with its ancestral viruses in bats in the counteraction of virus sensing. This work not only provides a new understanding of the abilities of MERS-CoV and closely related bat viruses to subvert virus sensing but also might prove useful in revealing new strategies for the development of vaccines and antivirals.
Functional Rectification of the Newly Described African Henipavirus Fusion Glycoprotein (Gh-M74a)
Authors: Pernet Olivier , Beaty Shannon , Lee Benhur
Published: February 2014

Recent evidence identified multiple
Henipavirus
species in Africa distinct from those in Southeast Asia and Australia. The reported fusion glycoprotein (F) sequence of the African Gh-M74a strain (GhV-F) is likely incorrect: a single base pair deletion near the N terminus results in multiple aberrancies. Rectifying this by adding single nucleotide insertions results in a GhV-F that now possesses a signal peptide, is efficiently cell surface expressed, exhibits syncytium formation when coexpressed with GhV-G protein, and mediates pseudotyped viral particle entry.
Epstein-Barr Virus Utilizes Ikaros in Regulating Its Latent-Lytic Switch in B Cells
Authors: Iempridee Tawin , Reusch Jessica A. , Riching Andrew , Johannsen Eric C. , Dovat Sinisa , Kenney Shannon C. , Mertz Janet E.
Published: February 2014

Ikaros is a zinc finger DNA-binding protein that regulates chromatin remodeling and the expression of genes involved in the cell cycle, apoptosis, and Notch signaling. It is a master regulator of lymphocyte differentiation and functions as a tumor suppressor in acute lymphoblastic leukemia. Nevertheless, no previous reports described effects of Ikaros on the life cycle of any human lymphotropic virus. Here, we demonstrate that full-length Ikaros (IK-1) functions as a major factor in the maintenance of viral latency in Epstein-Barr virus (EBV)-positive Burkitt's lymphoma Sal and MutuI cell lines. Either silencing of Ikaros expression by small hairpin RNA (shRNA) knockdown or ectopic expression of a non-DNA-binding isoform induced lytic gene expression. These effects synergized with other lytic inducers of EBV, including transforming growth factor β (TGF-β) and the hypoxia mimic desferrioxamine. Data from chromatin immunoprecipitation (ChIP)-quantitative PCR (qPCR) and ChIP-sequencing (ChIP-seq) analyses indicated that Ikaros did not bind to either of the EBV immediate early genes
BZLF1
and
BRLF1
. Rather, Ikaros affected the expression of Oct-2 and Bcl-6, other transcription factors that directly inhibit EBV reactivation and plasma cell differentiation, respectively. IK-1 also complexed with the EBV immediate early R protein in coimmunoprecipitation assays and partially colocalized with R within cells. The presence of R alleviated IK-1-mediated transcriptional repression, with IK-1 then cooperating with Z and R to enhance lytic gene expression. Thus, we conclude that Ikaros plays distinct roles at different stages of EBV's life cycle: it contributes to maintaining latency via indirect mechanisms, and it may also synergize with Z and R to enhance lytic replication through direct association with R and/or R-induced alterations in Ikaros' functional activities via cellular signaling pathways.

IMPORTANCE
This is the first report showing that the cellular protein Ikaros, a known master regulator of hematopoiesis and critical tumor suppressor in acute lymphoblastic leukemia, also plays important roles in the life cycle of Epstein-Barr virus in B cells.