bluuebunny/crossref_metadata_2025_split huggingface dataset:
Randomized, Double-Blind Crossover Study of Vaginal Microflora and Epithelium in Women Using a Tampon with a “Winged” Apertured Film Cover and a Commercial Tampon with a Nonwoven Fleece Cover
Authors: Chase David J. , Schenkel Berenike P. , Fahr Anne-Marie , Eigner Ulrich 
Published: February 2010

ABSTRACT

This study compared the safety of a new tampon with a four-winged apertured film cover over its nonwoven cover to improve leakage performance with that of a commercial tampon with a nonwoven cover only. Healthy women (evaluable,
n
= 69) were randomized to crossover between test and reference tampons in two consecutive menstrual cycles. Qualitative and quantitative analyses of vaginal cultures were conducted pre-, mid-, and postmenstrually for a broad panel of microorganisms, and colposcopy was performed. Similar to previous studies, prevalence and mean colony counts of the majority of microorganisms generally increased midmenstrually and returned or began to return postmenstrually. In contrast to most previous studies,
Lactobacillus
species remained at similar levels throughout the cycles with both tampons. Neither tampon was associated with clinically significant microbiological changes or abnormalities or with vaginal/cervical epithelial integrity changes on colposcopy. Microbiological and colposcopic evaluations indicate that the apertured film-covered tampon is safe.
Correction of Underquantification of Human Immunodeficiency Virus Type 1 Load with the Second Version of the Roche Cobas AmpliPrep/Cobas TaqMan Assay
Authors: De Bel A. , Marissens D. , Debaisieux L. , Liesnard C. , Van den Wijngaert S. , Lauwers S. , Piérard D. 
Published: February 2010

ABSTRACTInitial evaluations of the Cobas AmpliPrep/Cobas TaqMan human immunodeficiency virus type 1 (HIV-1) test (CAP/CTM) demonstrated good performance but, afterwards, reports about underquantification were published. We investigated whether the problem was solved with a second version of this assay, the Cobas AmpliPrep/Cobas TaqMan HIV-1 test, version 2.0 (CAP/CTM v2.0). The remaining plasma of 375 consecutive HIV-1 positive samples with a viral load of ≥4,000 copies/ml was collected in three laboratories. The samples were diluted and retested with our routine method Cobas AmpliPrep/Cobas Amplicor HIV-1 monitor test v1.5 in ultrasensitive mode (CAP/CA PHS), as well as with the CAP/CTM and CAP/CTM v2.0 tests. An absolute difference between the results of two methods of ≥0.71 log10copies/ml was defined as moderately discrepant, and an absolute difference of ≥0.93 log10copies/ml was defined as severely discrepant. In addition, criteria for considering the new methods equivalent to the routine method were formulated. (i) For CAP/CTM compared to CAP/CA PHS, 36 (9.5%) and 20 (5.3%) samples were, respectively, considered moderately and severely underquantified by CAP/CTM. The mean difference between CAP/CTM and CAP/CA PHS was −0.32 log10copies/ml. Eight of nineteen of the severely underquantified samples were from patients infected with HIV-1 subtype B strain. (ii) For CAP/CTM v2.0 compared to CAP/CA PHS, no sample was moderately or severely underquantified by CAP/CTM v2.0. A mean difference of 0.08 log10copies/ml was found with CAP/CTM v2.0 compared to CAP/CA PHS. The underquantification problem of the CAP/CTM kit was clearly demonstrated. The criteria for the equivalence of CAP/CTM v2.0 to the routine test CAP/CA PHS were fulfilled.
Development of a Multiplex Primer Extension Assay for Rapid Detection of
Salmonella
Isolates of Diverse Serotypes
Authors: Ben-Darif Elloulu , Jury Francine , De Pinna Elizabeth , Threlfall E. John , Bolton Frederick J. , Fox Andrew J. , Upton Mathew 
Published: February 2010

ABSTRACT

Food-borne salmonellosis is a major manifestation of gastrointestinal disease in humans across the globe. Accurate and rapid identification methods could positively impact the identification of isolates, enhance outbreak investigation, and aid infection control. The SNaPshot multiplex system is a primer extension-based method that enables multiplexing of single nucleotide polymorphisms (SNPs). Here the method has been developed for the identification of five
Salmonella
serotypes, commonly detected in the United Kingdom, based on serotype-specific SNPs identified in the multilocus sequence typing (MLST) database of
Salmonella enterica
. The SNPs, in genes
hemD
,
thrA
,
purE
, and
sucA
, acted as surrogate markers for
S. enterica
serovars Typhimurium, Enteritidis, Virchow, Infantis, and Braenderup. The multiplex primer extension assay (MPEA) was conducted in two separate panels and evaluated using 152
Salmonella enterica
isolates that were characterized by MLST. The MPEA was shown to be 100% specific and sensitive, within this collection of isolates. The MPEA is a sensitive and specific method for the identification and detection of
Salmonella
serotypes based upon SNPs seen in MLST data. The method can be applied in less than 6 h and has the potential to improve patient care and source tracing. The utility of the assay for identification of
Salmonella
serotypes directly from clinical specimens and food samples warrants further investigation.
Evaluation of Chromogenic Media for Detection of Methicillin-Resistant
Staphylococcus aureus
Authors: Malhotra-Kumar Surbhi , Abrahantes José Cortinas , Sabiiti Wilber , Lammens Christine , Vercauteren Greetje , Ieven Margareta , Molenberghs Geert , Aerts Marc , Goossens Herman 
Published: February 2010

ABSTRACT

Rapid laboratory diagnosis is critical for treating, managing, and preventing methicillin-resistant
Staphylococcus aureus
(MRSA) infections. We evaluated and compared the potential for MRSA detection of five chromogenic media,
Brilliance
MRSA agar (Oxoid), ChromID (bioMérieux), MRSA
Select
(Bio-Rad), CHROMagar (CHROMagar Microbiology), and BBL-CHROMagar (BD Diagnostics). Media were tested with log serial dilutions (10
0
to 10
6
CFU) of pure isolates of MRSA (
n
= 60), non-MRSA (
n
= 27), and defined mixtures thereof simulating clinical samples (
n
= 84). Further evaluations were done on pre-enriched nasal and groin screening swabs (
n
= 213) from 165 hospitalized patients. Randomized samples were spiral plated on each medium and independently scored by five investigators for characteristic colonies at 24 and 48 h of incubation. Confirmatory testing of up to five putative MRSA colonies recovered from each medium was done. The cumulative average sensitivity with isolates, mixtures, and clinical samples was the highest for
Brilliance
MRSA agar (97%) and similar for the other four media (≥92%). The cumulative average specificity was the highest for BBL-CHROMagar (99%), followed by MRSA
Select
(98%), CHROMagar (97%), ChromID (89%), and
Brilliance
MRSA agar (86%). All of the media detected MRSA at 10 and 1 CFU, although at these low loads, few MRSA samples harboring SCC
mec
type III or IV were misinterpreted as non-MRSA by investigators. False-positive results were mainly due to methicillin-resistant
S. epidermidis
. For an arbitrary MRSA prevalence of 5% and based on patient sample evaluations, the positive predictive values for BBL-CHROMagar and CHROMagar (∼84%) were the highest. The negative predictive values of all of the media were ≥92% for MRSA prevalences ranging from 5% to 30%. In conclusion, BBL-CHROMagar and CHROMagar gave the best overall results for detection of MRSA, irrespective of the sample concentration, investigator, or incubation period.
Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for Direct Bacterial Identification from Positive Blood Culture Pellets
Authors: Prod'hom Guy , Bizzini Alain , Durussel Christian , Bille Jacques , Greub Gilbert 
Published: February 2010

ABSTRACT
An ammonium chloride erythrocyte-lysing procedure was used to prepare a bacterial pellet from positive blood cultures for direct matrix-assisted laser desorption-ionization time of flight (MALDI-TOF) mass spectrometry analysis. Identification was obtained for 78.7% of the pellets tested. Moreover, 99% of the MALDI-TOF identifications were congruent at the species level when considering valid scores. This fast and accurate method is promising.
Performance Evaluation of the New Roche Cobas AmpliPrep/Cobas TaqMan HIV-1 Test Version 2.0 for Quantification of Human Immunodeficiency Virus Type 1 RNA
Authors: Pas Suzan , Rossen John W. A. , Schoener Daniel , Thamke Diana , Pettersson Annika , Babiel Reiner , Schutten Martin 
Published: February 2010

ABSTRACT

Despite FDA approval and CE marking of commercial tests, manufacturer-independent testing of the technical aspects of newly developed tests is important. To evaluate the analytical performance and explore the clinical applicability of the new Roche COBAS AmpliPrep COBAS TaqMan HIV-1 test, version 2.0 (CAP/CTM v2.0), platform comparison was performed with the Roche CAP/CTM test, version 2.0, the COBAS Amplicor HIV-1 Monitor Test, version 1.5 (CAP/CA v1.5), the COBAS AmpliPrep COBAS TaqMan HIV-1 Test (CAP/CTM v1.0), and the Abbott
m
2000 Real
Time
HIV-1 assay on panels and diagnostic samples. Specificity was tested for HIV-2 samples. Furthermore, samples from HIV-1-seropositive individuals with CAP/CA v1.5-measured viral loads below 50 HIV-1 RNA copies per ml (cp/ml) and replicates of HIV-1-seronegative plasma were tested in a checkerboard analysis. CAP/CTM v2.0 is HIV-1 specific, with broad genotype inclusivity and no serious underquantification of viral load relative to the other assays used. Low viral loads below the threshold of quantification for CAP/CA v1.5 are observed with CAP/CTM v2.0. A CAP/CTM v2.0-measured viral load of >50 copies/ml in these samples correlated with therapy failure. In conclusion, CAP/CTM v2.0 is an accurate and reliable test for HIV-1 viral load measurement relative to the other assays used with respect to specificity, sensitivity, and genotype inclusivity.
Catheter-Related Bacteremia Caused by
Staphylococcus pseudintermedius
Refractory to Antibiotic-Lock Therapy in a Hemophilic Child with Dog Exposure
Authors: Chuang Chia-Yunn , Yang Yung-Li , Hsueh Po-Ren , Lee Ping-Ing 
Published: February 2010

ABSTRACT

We describe a case of catheter-related bacteremia due to
Staphylococcus pseudintermedius
in a child with dog exposure. The organism was confirmed as
S
.
pseudintermedius
based on 16S rRNA gene sequence analysis and positive PCR-restriction fragment length polymorphism of the
pta
gene.
Decreased Susceptibility to Teicoplanin and Vancomycin in Coagulase-Negative Staphylococci Isolated from Orthopedic-Device-Associated Infections
Authors: Cremniter Julie , Slassi Asma , Quincampoix Jean-Charles , Sivadon-Tardy Valérie , Bauer Thomas , Porcher Raphaël , Lortat-Jacob Alain , Piriou Philippe , Judet Thierry , Herrmann Jean-Louis , Gaillard Jean-Louis , Rottman Martin 
Published: February 2010

ABSTRACT

We studied 315 coagulase-negative
Staphylococcus
strains recovered prospectively during 240 surgical procedures (206 subjects) from proven or suspected device-associated bone and joint infections. Sixteen strains (5.1%) had decreased susceptibility to glycopeptides: 15 (12
S. epidermidis
strains, 2
S. capitis
strains, and 1
S. haemolyticus
strain) to teicoplanin alone (MIC of 16 mg/liter,
n
= 9; MIC of 32 mg/liter,
n
= 6) and one (
S. epidermidis
) to both teicoplanin and vancomycin (MIC, 16 and 8 mg/liter, respectively). Decreased susceptibility to teicoplanin was more prevalent in “infecting” strains (i.e., strains recovered from ≥2 distinct intraoperative samples) than in “contaminants” (i.e., strains not fulfilling this criterion) (8.1% [12/149] versus 2.4% [4/166], respectively [
P
= 0.022]). One hundred percent (13/13) of
S. epidermidis
strains with decreased susceptibility to teicoplanin were resistant to methicillin (versus 112/173 [64.7%] for
S. epidermidis
strains susceptible to teicoplanin;
P
= 0.021).
Frequent Detection of Noroviruses and Sapoviruses in Swine and High Genetic Diversity of Porcine Sapovirus in Japan during Fiscal Year 2008
Authors: Nakamura Kazuya , Saga Yumiko , Iwai Masae , Obara Mayumi , Horimoto Eiji , Hasegawa Sumiyo , Kurata Takeshi , Okumura Hiraku , Nagoshi Masataka , Takizawa Takenori 
Published: February 2010

ABSTRACT
A molecular biological survey on porcine norovirus (NoV) and sapovirus (SaV) was conducted in Toyama Prefecture, Japan, during fiscal year 2008. Both NoV and SaV were detected from swine fecal samples throughout the surveillance period, indicating that these viruses were circulating in this region. NoV strains detected in this study belonged to three genotypes that are known as typical swine NoVs. Although human NoVs were occasionally detected, it was unclear whether they replicated in pigs. As for SaV, genogroup VII (GVII) and other divergent genogroups were identified in addition to the dominant genogroup, GIII, which is the prototypic porcine SaV. In addition, 3 strains genetically related to human SaV were detected. Two of these 3 strains were closely related to human SaV GV. Our study showed that genetic diversification of porcine SaV is currently progressing in the swine population.
Application of an Improved Enzyme-Linked Immunosorbent Assay Method for Serological Diagnosis of Canine Leishmaniasis
Authors: Santarém Nuno , Silvestre Ricardo , Cardoso Luís , Schallig Henk , Reed Steven G. , Cordeiro-da-Silva Anabela 
Published: February 2010

ABSTRACT

Accurate diagnosis of canine leishmaniasis (CanL) is essential toward a more efficient control of this zoonosis, but it remains problematic due to the high incidence of asymptomatic infections. In this study, we present data on the development of enzyme-linked immunosorbent assay (ELISA)-based techniques for the detection of antibodies against the recombinant protein
Leishmania infantum
cytosolic tryparedoxin peroxidase (
Li
cTXNPx) and a comparison of the results with those employing soluble
Leishmania
antigens from promastigote or amastigote forms and the homologue recombinant protein
L. infantum
mitochondrial TXNPx (
Li
mTXNPx). Moreover, we offer an evaluation of the diagnostic potential of rK39 for CanL in the Portuguese canine population and propose an improvement to the existing ELISA-based serological techniques by combining the
Li
cTXNPx and rK39 antigens as a
Leishmania
antigen mixture (LAM). The data demonstrated that ELISAs based on soluble promastigote or amastigote antigens had generally higher levels of sensitivity for detection of antibodies in symptomatic or asymptomatic dogs than for detection of those against isolated recombinant proteins. Nevertheless, the specificities were found to be similar for all target antigens used. Importantly, the LAM-ELISA methodology improved the overall sensitivity, maintaining a high overall level of specificity. In addition, it was demonstrated that the detection of anti-LAM IgG2 can increase the accuracy of the serological diagnosis. Overall, the obtained results showed that the strategy of combining two well-defined
Leishmania
antigens,
Li
cTXNPx and rK39, proved to be a sensitive and specific improvement to current serological diagnosis of CanL, being a useful tool for the detection of both clinical and subclinical forms of canine
Leishmania
infection.
Interaction of a Host Protein with Core Complexes of Bacteriophage Φ6 To Control Transcription
Authors: Qiao Xueying , Sun Yang , Qiao Jian , Mindich Leonard 
Published: February 2010

ABSTRACTBacteriophages of the familyCystoviridaehave genomes consisting of three double-stranded RNA (dsRNA) segments, L, S, and M, packaged within a polyhedral capsid along with RNA polymerase. Transcription of genomic segment L is activated by the interaction of host protein YajQ with the capsid structure. Segment L codes for the proteins of the inner capsid, which are expressed early in infection. Green fluorescent protein (GFP) fusions with YajQ produce uniform fluorescence in uninfected cells and in cells infected with viruses not dependent on YajQ. Punctate fluorescence develops when cells are infected with YajQ-dependent viruses. It appears that the host protein binds to the infecting particles and remains with them during the entire infection period.
Reciprocal Regulation of AKT and MAP Kinase Dictates Virus-Host Cell Fusion
Authors: Sharma Nishi R. , Mani Prashant , Nandwani Neha , Mishra Rajakishore , Rana Ajay , Sarkar Debi P. 
Published: February 2010

ABSTRACT

Viruses of the
Paramyxoviridae
family bind to their host cells by using hemagglutinin-neuraminidase (HN), which enhances fusion protein (F)-mediated membrane fusion. Although respiratory syncytial virus and parainfluenza virus 5 of this family are suggested to trigger host cell signaling during infection, the virus-induced intracellular signals dictating virus-cell fusion await elucidation. Using an F- or HN-F-containing reconstituted envelope of Sendai virus, another paramyxovirus, we revealed the role and regulation of AKT1 and Raf/MEK/ERK cascades during viral fusion with liver cells. Our observation that extracellular signal-regulated kinase (ERK) activation promotes viral fusion via ezrin-mediated cytoskeletal rearrangements, whereas AKT1 attenuates fusion by promoting phosphorylation of F protein, indicates a counteractive regulation of viral fusion by reciprocal activation of AKT1 and mitogen-activated protein kinase (MAPK) cascades, establishing a novel conceptual framework for a therapeutic strategy.
Familial Relationships in Hyperthermo- and Acidophilic Archaeal Viruses
Authors: Happonen Lotta Johanna , Redder Peter , Peng Xu , Reigstad Laila Johanne , Prangishvili David , Butcher Sarah Jane 
Published: February 2010

ABSTRACT

Archaea often live in extreme, harsh environments such as acidic hot springs and hypersaline waters. To date, only two icosahedrally symmetric, membrane-containing archaeal viruses, SH1 and
Sulfolobus
turreted icosahedral virus (STIV), have been described in detail. We report the sequence and three-dimensional structure of a third such virus isolated from a hyperthermoacidophilic crenarchaeon,
Sulfolobus
strain G4ST-2. Characterization of this new isolate revealed it to be similar to STIV on the levels of genome and structural organization. The genome organization indicates that these two viruses have diverged from a common ancestor. Interestingly, the prominent surface turrets of the two viruses are strikingly different. By sequencing and mass spectrometry, we mapped several large insertions and deletions in the known structural proteins that could account for these differences and showed that both viruses can infect the same host. A combination of genomic and proteomic analyses revealed important new insights into the structural organization of these viruses and added to our limited knowledge of archaeal virus life cycles and host-cell interactions.
Mechanistic Consequences of hnRNP C Binding to Both RNA Termini of Poliovirus Negative-Strand RNA Intermediates
Authors: Ertel Kenneth J. , Brunner Jo Ellen , Semler Bert L. 
Published: February 2010

ABSTRACT

The poliovirus 3′ noncoding region (3′ NCR) is necessary for efficient virus replication. A poliovirus mutant, PVΔ3′NCR, with a deletion of the entire 3′ NCR, yielded a virus that was capable of synthesizing viral RNA, albeit with a replication defect caused by deficient positive-strand RNA synthesis compared to wild-type virus. We detected multiple ribonucleoprotein (RNP) complexes in extracts from poliovirus-infected HeLa cells formed with a probe corresponding to the 5′ end of poliovirus negative-strand RNA (the complement of the genomic 3′ NCR), and the levels of these RNP complexes increased during the course of viral infection. Previous studies have identified RNP complexes formed with the 3′ end of poliovirus negative-strand RNA, including one that contains a 36-kDa protein later identified as heterogeneous nuclear ribonucleoprotein C (hnRNP C). We report here that the 5′ end of poliovirus negative-strand RNA is capable of interacting with endogenous hnRNP C, as well as with poliovirus nonstructural proteins. Further, we demonstrate that the addition of recombinant purified hnRNP C proteins can stimulate virus RNA synthesis
in vitro
and that depletion of hnRNP C proteins in cultured cells results in decreased virus yields and a correspondingly diminished accumulation of positive-strand RNAs. We propose that the association of hnRNP C with poliovirus negative-strand termini acts to stabilize or otherwise promote efficient positive-strand RNA synthesis.
Herpes Simplex Virus 1 Regulatory Protein ICP27 Undergoes a Head-to-Tail Intramolecular Interaction
Authors: Hernandez Felicia P. , Sandri-Goldin Rozanne M. 
Published: February 2010

ABSTRACT
Herpes simplex virus type 1 (HSV-1) regulatory protein ICP27 is a multifunction functional protein that interacts with many cellular proteins. A number of the proteins with which ICP27 interacts require that both the N and C termini of ICP27 are intact. These include RNA polymerase II, TAP/NXF1, and Hsc70. We tested the possibility that the N and C termini of ICP27 could undergo a head-to-tail intramolecular interaction that exists in open and closed configurations for different binding partners. Here, we show by bimolecular fluorescence complementation (BiFC) assays and fluorescence resonance energy transfer (FRET) by acceptor photobleaching that ICP27 undergoes a head-to-tail intramolecular interaction but not head-to-tail or tail-to-tail intermolecular interactions. Substitution mutations in the N or C termini showed that the leucine-rich region (LRR) in the N terminus and the zinc finger-like region in the C terminus must be intact for intramolecular interactions. A recombinant virus, vNC-Venus-ICP27, was constructed, and this virus was severely impaired for virus replication. The expression of NC-Venus-ICP27 protein was delayed compared to ICP27 expression in wild-type HSV-1 infection, but NC-Venus-ICP27 was abundantly expressed at late times of infection. Because the renaturation of the Venus fluorescent protein results in a covalent bonding of the two halves of the Venus molecule, the head-to-tail interaction of NC-Venus-ICP27 locks ICP27 in a closed configuration. We suggest that the population of locked ICP27 molecules is not able to undergo further protein-protein interactions.
Inhibition of Human Natural Killer Cell Activity by Influenza Virions and Hemagglutinin
Authors: Mao Huawei , Tu Wenwei , Liu Yinping , Qin Gang , Zheng Jian , Chan Ping-Lung , Lam Kwok-Tai , Peiris J. S. Malik , Lau Yu-Lung 
Published: February 2010

ABSTRACTNatural killer (NK) cells keep viral infections under control at the early phase by directly killing infected cells. Influenza is an acute contagious respiratory viral disease transmitted from host-to-host in the first few days of infection. The evasion of host innate immune defenses including NK cells is important for its success as a viral pathogen of humans and animals. NK cells encounter influenza virus within the microenvironment of infected cells. It therefore is important to investigate the direct effects of influenza virus on NK cell activity. Recently we demonstrated that influenza virus directly infects human NK cells and induces cell apoptosis to counter their function (H. Mao, W. Tu, G. Qin, H. K. W. Law, S. F. Sia, P.-L. Chan, Y. Liu, K.-T. Lam, J. Zheng, M. Peiris, and Y.-L. Lau, J. Virol. 83:9215-9222, 2009). Here, we further demonstrated that both the intact influenza virion and free hemagglutinin protein inhibited the cytotoxicity of fresh and interleukin-2 (IL-2)-activated primary human NK cells. Hemagglutinin bound and internalized into NK cells via the sialic acids. This interaction did not decrease NKp46 expression but caused the downregulation of the ζ chain through the lysosomal pathway, which caused the decrease of NK cell cytotoxicity mediated by NKp46 and NKp30. The underlying dysregulation of the signaling pathway involved ζ chain downregulation, leading to decreased Syk and ERK activation and granule exocytosis upon target cell stimulation, finally causing reduced cytotoxicity. These findings suggest that influenza virus developed a novel strategy to evade NK cell innate immune defense that is likely to facilitate viral transmission and also contribute to virus pathogenesis.