bluuebunny/crossref_metadata_2025_split huggingface dataset:
Persistence of Livestock-Associated Methicillin-Resistant Staphylococcus aureus in Field Workers after Short-Term Occupational Exposure to Pigs and Veal Calves
Authors: van Cleef Brigitte A. G. L. , Graveland Haitske , Haenen Anja P. J. , van de Giessen Arjen W. , Heederik Dick , Wagenaar Jaap A. , Kluytmans Jan A. J. W. 
Published: January 2011

ABSTRACT

The prevalence of methicillin-resistant
Staphylococcus aureus
(MRSA) carriage in pig and veal calf farmers in the Netherlands is estimated at 25 to 35%. However, no information is available about MRSA carriage in humans after short-term occupational exposure to pigs or veal calves. This study examines the prevalence and duration of MRSA acquisition after short-term intensive exposure to pigs or veal calves for persons not exposed to livestock on a daily basis. The study was performed with field workers who took samples from the animals or the animal houses in studies on MRSA prevalence in pig and veal farms. They were tested for MRSA by taking nasal samples before, directly after, and 24 h after they visited the farms. There were 199 sampling moments from visits to 118 MRSA-positive farms. Thirty-four of these visits (17%) resulted in the acquisition of MRSA. Thirty-one persons (94%) appeared negative again after 24 h. There were 62 visits to 34 MRSA-negative farms; none of the field workers acquired MRSA during these visits. Except for that from one person, all
spa
types found in the field workers were identical to those found in the animals or in the dust in animal houses and belonged to the livestock-associated clone. In conclusion, MRSA is frequently present after short-term occupational exposure, but in most cases the strain is lost again after 24 h.
Cellulosimicrobium funkei: First Report of Infection in a Nonimmunocompromised Patient and Useful Phenotypic Tests for Differentiation from Cellulosimicrobium cellulans and Cellulosimicrobium terreum
Authors: Petkar Hawabibee , Li Anthony , Bunce Nicholas , Duffy Kim , Malnick Henry , Shah Jayesh J. 
Published: January 2011

ABSTRACT

Cellulosimicrobium funkei
is a rare, opportunistic pathogen. We describe a case of bacteremia and possibly prosthetic valve endocarditis by this organism in a nonimmunocompromised patient. Useful phenotypic tests for differentiating
C. funkei
from
Cellulosimicrobium cellulans
and
Cellulosimicrobium terreum
include motility, raffinose fermentation, glycogen,
d
-xylose, and methyl-α-
d
-glucopyranoside assimilation, and growth at 35°C.
Interpretation of Real-Time PCR Results for Hepatitis C Virus RNA When Viral Load Is Below Quantification Limits
Authors: Laperche Syria , Bouchardeau Françoise , André-Garnier Elisabeth , Thibault Vincent , Roque-Afonso Anne-Marie , Trimoulet Pascale , Colimon Ronald , Duverlie Gilles , Leguillou-Guillemette Hélène , Lunel Françoise , Bouvier-Alias Magali , Pawlotsky Jean-Michel , Henquell Cécile , Schvoerer Evelyne , Stoll-Keller Françoise , Chaix Marie-Laure , Branger Michel , Gaudy-Graffin Catherine , Rosenberg Arielle R. , Pozzetto Bruno , Vallet Sophie , Baazia Yazid , Izopet Jacques , Lefrère Jean-Jacques 
Published: January 2011

ABSTRACT
Hepatitis C virus RNA quantification results obtained in 18 laboratories using real-time PCR methods with 10 negative samples and 22 sample dilutions (viral loads of 0.5 to 500 IU/ml) showed a score of correct results of up to 93.5%. However, 55.6% of the laboratories did not follow the recommendations for the interpretation of their results, leading to ambiguous conclusions.
Human and Swine Hosts Share Vancomycin-Resistant Enterococcus faecium CC17 and CC5 and Enterococcus faecalis CC2 Clonal Clusters Harboring Tn
1546
on Indistinguishable Plasmids
Authors: Freitas Ana R. , Coque Teresa M. , Novais Carla , Hammerum Anette M. , Lester Camilla H. , Zervos Marcus J. , Donabedian Susan , Jensen Lars B. , Francia Maria Victoria , Baquero Fernando , Peixe Luísa 
Published: January 2011

ABSTRACT

VRE isolates from pigs (
n
= 29) and healthy persons (
n
= 12) recovered during wide surveillance studies performed in Portugal, Denmark, Spain, Switzerland, and the United States (1995 to 2008) were compared with outbreak/prevalent VRE clinical strains (
n
= 190; 23 countries; 1986 to 2009). Thirty clonally related
Enterococcus faecium
clonal complex 5 (CC5) isolates (17 sequence type 6 [ST6], 6 ST5, 5 ST185, 1 ST147, and 1 ST493) were obtained from feces of swine and healthy humans. This collection included isolates widespread among pigs of European Union (EU) countries since the mid-1990s. Each ST comprised isolates showing similar pulsed-field gel electrophoresis (PFGE) patterns (≤6 bands difference; >82% similarity). Some CC5 PFGE subtype strains from swine were indistinguishable from hospital vancomycin-resistant enterococci (VRE) causing infections. A truncated variant of Tn
1546
(encoding resistance to vancomycin) and
tcrB
(coding for resistance to copper) were consistently located on 150- to 190-kb plasmids (rep
pLG1
).
E. faecium
CC17 (ST132) isolates from pig manure and two clinical samples showed identical PFGE profiles and contained a 60-kb mosaic plasmid (rep
Inc18
plus rep
pRUM
) carrying diverse Tn
1546
-IS
1216
variants. The only
Enterococcus faecalis
isolate obtained from pigs (CC2-ST6) corresponded to a multidrug-resistant clone widely disseminated in hospitals in Italy, Portugal, and Spain, and both animal and human isolates harbored an indistinguishable 100-kb mosaic plasmid (rep
pRE25
plus rep
pCF10
) containing the whole Tn
1546
backbone. The results indicate a current intra- and international spread of
E. faecium
and
E. faecalis
clones and their plasmids among swine and humans.
Emergence in Spain of a Multidrug-Resistant Enterobacter cloacae Clinical Isolate Producing SFO-1 Extended-Spectrum β-Lactamase
Authors: Fernández Ana , Pereira María José , Suárez José Manuel , Poza Margarita , Treviño Mercedes , Villalón Pilar , Sáez-Nieto Juan Antonio , Regueiro Benito José , Villanueva Rosa , Bou Germán 
Published: January 2011

ABSTRACT

Between February 2006 and October 2009, 38 patients in different wards at the A Coruña University Hospital (northwest Spain) were either infected with or colonized by an epidemic, multidrug-resistant (MDR), and extended-spectrum-β-lactamase (ESBL)-producing strain of
Enterobacter cloacae
(EbSF), which was susceptible only to carbapenems. Semiautomated repetitive extragenic palindromic sequence-based PCR (rep-PCR) and pulsed-field gel electrophoresis (PFGE) analysis revealed that all of the
E. cloacae
isolates belonged to the same clone. Cloning and sequencing enabled the detection of the SFO-1 ESBL in the epidemic strain and the description of its genetic environment. The presence of the
ampR
gene was detected upstream of
bla
SFO-1
, and two complete sequences of IS
26
surrounding
ampR
and
ampA
were detected. These IS
26
sequences are bordered by complete left and right inverted repeats (IRL and IRR, respectively), which suggested that they were functional. The whole segment flanked by two IS
26
copies may be considered a putative large composite transposon. A gene coding for aminoglycoside acetyltransferase (gentamicin resistance gene [
aac3
]) was found downstream of the 3′ IS
26
. Despite the implementation of strict infection control measures, strain EbSF spread through different areas of the hospital. A case-control study was performed to assess risk factors for EbSF acquisition. A multivariate analysis revealed that the prior administration of β-lactam antibiotics, chronic renal failure, tracheostomy, and prior hospitalization were statistically associated with SFO-1-producing
E. cloacae
acquisition. This study describes for the first time an outbreak in which an SFO-1-producing
E. cloacae
strain was involved. Note that so far, this β-lactamase has previously been isolated in only a single case of
E. cloacae
infection in Japan.
Ribosomal and Mitochondrial DNA Target for Real-Time PCR Diagnosis of Invasive Aspergillosis
Authors: Millon L. , Grenouillet F. , Legrand F. , Loewert S. , Bellanger A. P. , Gbaguidi-Haore H. , Scherer E. , Henon T. , Rohrlich P. , Deconinck E. 
Published: January 2011

ABSTRACT

The aim of the present study was to assess the diagnostic efficacy of a combination of two quantitative
Aspergillus
PCR assays, targeting a mitochondrial and a ribosomal target, in patients with risk factors for invasive aspergillosis (IA) and positive galactomannan (GM) antigen. Forty-four patients with hematological malignancies and risk factors for IA according to revised European Organization for Research on Treatment of Cancer and the Mycoses Study Group criteria (EORTC/MSG) criteria and presenting at least two sequential GM-positive sera were included in the study. Mitochondrial PCR was carried out prospectively on all GM-positive serum samples. Ribosomal PCR was carried out retrospectively on frozen stored sera. The sensitivities of mitochondrial and ribosomal PCRs were 58% and 50%, respectively. The diagnostic test performance was improved by using a combination of both PCR assays and by considering a patient PCR positive when at least two positive results were obtained. The sensitivity, specificity, and positive and negative likelihood ratios were 65%, 94%, and 11.8 and 0.37, respectively. A significant association between fatal outcome at 90 days and positive results of ribosomal PCR assays was observed (adjusted hazard ratio = 8.2; 95% confidence interval [CI] = 1.0 to 65.8;
P
= 0.048). Our results showed that the combination of two PCR assays targeting mitochondrial and ribosomal
Aspergillus
DNA improves the sensitivity of PCR in the diagnosis of IA in hematological patients with risk factors and positive GM results. This study also confirms that a positive PCR result is associated with a poor prognosis in these patients and should lead to specific antifungal therapy being introduced immediately.
Optimizing a Candida Biofilm Microtiter Plate Model for Measurement of Antifungal Susceptibility by Tetrazolium Salt Assay
Authors: Nett Jeniel E. , Cain Michael T. , Crawford Kyler , Andes David R. 
Published: January 2011

ABSTRACT

Candida
spp. infect medical devices, such as venous and urinary catheters, by adhering to the surface and forming a community of drug-resistant cells surrounded by a matrix. The ability to measure drug activity during this biofilm mode of growth is of interest for the investigation of resistance mechanisms and novel antifungal therapies. The tetrazolium salt (XTT) reduction assay is the test most commonly used to estimate viable biofilm growth and to examine the impact of biofilm therapies. The primary goal of the current experiments was to identify assay variables that affect the XTT assay result in order to improve assay reproducibility, sensitivity, and throughput for the study of antifungal activity. The species used in the current studies included
Candida albicans
,
C. parapsilosis
, and
C. glabrata
. The assay variables that were studied included the impact of culture conditions, the duration of biofilm growth, the timing and frequency of drug administration, the XTT source and concentration, and the duration of XTT incubation. The conditions that impacted the assay readout and altered assay sensitivity included the duration of biofilm growth, the frequency of drug dosing, and the duration of XTT incubation. Several factors were found to reduce time and assay expense, including the elimination of washing steps, the shortening of incubation times, and the use of lower XTT concentrations. A description of assay pitfalls and troubleshooting is included. Recognition of these technical variables should allow investigators to better design reproducible biofilm therapeutic studies.
Herpes Simplex Virus Requires VP11/12 To Activate Src Family Kinase-Phosphoinositide 3-Kinase-Akt Signaling
Authors: Wagner Melany J. , Smiley James R. 
Published: January 2011

ABSTRACTWe previously showed that the herpes simplex virus 1 (HSV-1) tegument protein VP11/12 activates the lymphocyte-specific Src family kinase (SFK) Lck and is tyrosine phosphorylated in an Lck-dependent manner during T cell infection. We now extend these findings to show that ectopic expression of Lck induces robust tyrosine phosphorylation of VP11/12 in Vero cells, strongly suggesting that VP11/12 participates in an Lck-mediated signaling pathway as a substrate of Lck or a kinase activated by Lck. We sought to elucidate signaling events downstream of VP11/12-SFK interactions. SFKs lie upstream of the canonical phosphoinositide 3-kinase (PI3K)-Akt pathway in signaling emanating from immune receptors, growth factor receptors, and polyomavirus middle T antigen. Here, we show that VP11/12 is required for virus-induced activation of PI3K-Akt signaling in HSV-infected Jurkat T cells and primary fibroblasts. VP11/12 interacts with PI3K or PI3K signaling complexes during infection, suggesting that VP11/12 activates PI3K directly. SFK activity is required for tyrosine phosphorylation of VP11/12, VP11/12-PI3K interactions, and Akt activation in infected fibroblasts, suggesting that SFK-dependent phosphorylation of VP11/12 is required for interactions with downstream signaling effectors. Akt controls many biological functions, including cell survival, cell motility, and translation, but it is currently unclear which Akt targets are modulated by VP11/12 during infection. Although the Akt target mTORC1 is activated during HSV-1 infection, VP11/12 is not required for this effect, implying that one or more additional viral proteins regulate this pathway. Further studies are therefore required to determine which Akt targets and associated biological functions are uniquely modulated by VP11/12.
Successful Propagation of Flavivirus Infectious cDNAs by a Novel Method To Reduce the Cryptic Bacterial Promoter Activity of Virus Genomes
Authors: Pu Szu-Yuan , Wu Ren-Huang , Yang Chi-Chen , Jao Tzu-Ming , Tsai Ming-Han , Wang Jing-Chyi , Lin Hui-Mei , Chao Yu-Sheng , Yueh Andrew 
Published: January 2011

ABSTRACT

Reverse genetics is a powerful tool to study single-stranded RNA viruses. Despite tremendous efforts having been made to improve the methodology for constructing flavivirus cDNAs, the cause of toxicity of flavivirus cDNAs in bacteria remains unknown. Here we performed mutational analysis studies to identify
Escherichia coli
promoter (ECP) sequences within nucleotides (nt) 1 to 3000 of the dengue virus type 2 (DENV2) and Japanese encephalitis virus (JEV) genomes. Eight and four active ECPs were demonstrated within nt 1 to 3000 of the DENV2 and JEV genomes, respectively, using fusion constructs containing DENV2 or JEV segments and empty vector reporter gene
Renilla
luciferase. Full-length DENV2 and JEV cDNAs were obtained by inserting mutations reducing their ECP activity in bacteria without altering amino acid sequences. A severe cytopathic effect occurred when BHK21 cells were transfected with
in vitro
-transcribed RNAs from either a DENV2 cDNA clone with multiple silent mutations within the prM-E-NS1 region of dengue genome or a JEV cDNA clone with an A-to-C mutation at nt 90 of the JEV genome. The virions derived from the DENV2 or JEV cDNA clone exhibited infectivities similar to those of their parental viruses in C6/36 and BHK21 cells. A
cis
-acting element essential for virus replication was revealed by introducing silent mutations into the central portion (nt 160 to 243) of the core gene of DENV2 infectious cDNA or a subgenomic DENV2 replicon clone. This novel strategy of constructing DENV2 and JEV infectious clones could be applied to other flaviviruses or pathogenic RNA viruses to facilitate research in virology, viral pathogenesis, and vaccine development.
Theiler's Virus L* Protein Is Targeted to the Mitochondrial Outer Membrane
Authors: Sorgeloos Frédéric , Vertommen Didier , Rider Mark H. , Michiels Thomas 
Published: January 2011

ABSTRACTThe L* protein encoded by Theiler's murine encephalomyelitis virus (TMEV) is a unique example of a picornaviral protein encoded by an alternative open reading frame. This protein is an important determinant of TMEV persistence in the mouse central nervous system. We showed that in infected cells, L* is partitioned between the cytosol and the mitochondria. In mitochondria, L* is anchored in the outer membrane and exposed to the cytosol. The targeting of L* to mitochondria is independent of other viral components and likely depends on a conformational signal. L* targeting to mitochondria might involve chaperones of the Hsp70 family, as these proteins are shown to interact.
Polyomavirus Middle T-Antigen Is a Transmembrane Protein That Binds Signaling Proteins in Discrete Subcellular Membrane Sites
Authors: Zhou Alice Y. , Ichaso Natalia , Adamarek Adam , Zila Vojtech , Forstova Jitka , Dibb Nicholas J. , Dilworth Stephen M. 
Published: January 2011

ABSTRACT
Murine polyomavirus middle T-antigen (MT) induces tumors by mimicking an activated growth factor receptor. An essential component of this action is a 22-amino-acid hydrophobic region close to the C terminus which locates MT to cell membranes. Here, we demonstrate that this sequence is a transmembrane domain (TMD) by showing that a hemagglutinin (HA) tag added to the MT C terminus is exposed on the outside of the cells, with the N terminus inside. To determine whether this MT TMD is inserted into the endoplasmic reticulum (ER) membrane, we added the ER retention signal KDEL to the MT C terminus (MTKDEL). This mutant protein locates only in the ER, demonstrating that MT does insert into membranes solely at this location. In addition, this ER-located MT failed to transform. Examination of the binding proteins associated with the MTKDEL protein demonstrated that it associates with PP2A and c-Src but fails to interact with ShcA, phosphatidylinositol 3-kinase (PI3K), and phospholipase C-γ1 (PLC-γ1), despite being tyrosine phosphorylated. Additional mutant and antibody studies show that MT binding to PP2A is probably required for MT to efficiently exit the ER and migrate to the plasma membrane though the TMD also plays a role in this relocation. Overall, these data, together with previous publications, illustrate that MT associates with signaling proteins at different sites in its maturation pathway. MT binds to PP2A in the cytoplasm, to c-Src at the endoplasmic reticulum, and to ShcA, PI3K, and PLC-γ1 at subsequent locations en route to the plasma membrane.
Vaccination against Seasonal Influenza A/H3N2 Virus Reduces the Induction of Heterosubtypic Immunity against Influenza A/H5N1 Virus Infection in Ferrets
Authors: Bodewes R. , Kreijtz J. H. C. M. , Geelhoed-Mieras M. M. , van Amerongen G. , Verburgh R. J. , van Trierum S. E. , Kuiken T. , Fouchier R. A. M. , Osterhaus A. D. M. E. , Rimmelzwaan G. F. 
Published: January 2011

ABSTRACTInfection with seasonal influenza viruses induces a certain extent of protective immunity against potentially pandemic viruses of novel subtypes, also known as heterosubtypic immunity. Here we demonstrate that infection with a recent influenza A/H3N2 virus strain induces robust protection in ferrets against infection with a highly pathogenic avian influenza virus of the H5N1 subtype. Prior H3N2 virus infection reduced H5N1 virus replication in the upper respiratory tract, as well as clinical signs, mortality, and histopathological changes associated with virus replication in the brain. This protective immunity correlated with the induction of T cells that cross-reacted with H5N1 viral antigen. We also demonstrated that prior vaccination against influenza A/H3N2 virus reduced the induction of heterosubtypic immunity otherwise induced by infection with the influenza A/H3N2 virus. The implications of these findings are discussed in the context of vaccination strategies and vaccine development aiming at the induction of immunity to pandemic influenza.
Participation of Vaccinia Virus L2 Protein in the Formation of Crescent Membranes and Immature Virions
Authors: Maruri-Avidal Liliana , Domi Arban , Weisberg Andrea S. , Moss Bernard 
Published: January 2011

ABSTRACT
Morphogenesis of vaccinia virus begins with the appearance of crescent-shaped membrane precursors of immature virions in cytoplasmic factories. During the initial characterization of the product of the L2R reading frame, we discovered that it plays an important role in crescent formation. The L2 protein was expressed early in infection and was associated with the detergent-soluble membrane fraction of mature virions, consistent with two potential membrane-spanning domains. All chordopoxviruses have L2 homologs, suggesting an important function. Indeed, we were unable to isolate an infectious L2R deletion mutant. Consequently, we constructed an inducible mutant with a conditional lethal phenotype. When L2 expression was repressed, proteolytic processing of the major core proteins and the A17 protein, which is an essential component of the immature virion membrane, failed to occur, suggesting an early block in viral morphogenesis. At 8 h after infection in the presence of inducer, immature and mature virions were abundantly seen by electron microscopy. In contrast, those structures were rare in the absence of inducer and were replaced by large, dense aggregates of viroplasm. A minority of these aggregates had short spicule-coated membranes, which resembled the beginnings of crescent formation, at their periphery. These short membrane segments at the edge of the dense viroplasm increased in number at later times, and some immature virions were seen. Although the L2 protein was not detected under nonpermissive conditions, minute amounts could account for stunted and delayed viral membrane formation. These findings suggested that L2 is required for the formation or elongation of crescent membranes.
Contribution of smoking-related and alcohol-related deaths to the gender gap in mortality: evidence from 30 European countries: Table 1
Authors: McCartney Gerry , Mahmood Lamia , Leyland Alastair H , Batty G David , Hunt Kate 
Published: January 2011

BackgroundWomen now outlive men throughout the globe, a mortality advantage that is very established in developed European countries. Debate continues about the causes of the gender gap, although smoking is known to have been a major contributor to the difference in the past.ObjectivesTo compare the magnitude of the gender gap in all-cause mortality in 30 European countries and assess the contribution of smoking-related and alcohol-related deaths.MethodsData on all-cause mortality, smoking-related mortality and alcohol-related mortality for 30 European countries were extracted from the World Health Organization Health for All database for the year closest to 2005. Rates were standardised by the direct method using the European population standard and were for all age groups. The proportion of the gender gap in all-cause mortality attributable to smoking-related and alcohol-related deaths was then calculated.ResultsThere was considerable variation in the magnitude of the male ‘excess’ of all-cause mortality across Europe, ranging from 188 per 100 000 per year in Iceland to 942 per 100 000 per year in Ukraine. Smoking-related deaths accounted for around 40% to 60% of the gender gap, while alcohol-related mortality typically accounted for 20% to 30% of the gender gap in Eastern Europe and 10% to 20% elsewhere in Europe.ConclusionsSmoking continues to be the most important cause of gender differences in mortality across Europe, but its importance as an explanation for this difference is often overshadowed by presumptions about other explanations. Changes in smoking patterns by gender suggest that the gender gap in mortality will diminish in the coming decades.