bluuebunny/crossref_metadata_2025_split huggingface dataset:
Antibodies against C-Reactive Protein Cross-React with 60-Kilodalton Heat Shock Proteins
Authors: Udvarnoki Katalin , Cervenak László , Uray Katalin , Hudecz Ferenc , Kacskovics Imre , Spallek Ralf , Singh Mahavir , Füst George , Prohászka Zoltán 
Published: February 2007

ABSTRACTC-reactive protein (CRP) is an acute-phase reactant frequently used in histochemistry as a marker of ongoing inflammation. Furthermore, CRP is a powerful biomarker for the prediction of coronary artery disease risk. Heat-shock protein 60 (Hsp60) and CRP are complement-activating molecules, and the effect of their interactions on the regulation of complement activation was studied. However, during the first experiments, we learned that polyclonal anti-CRP antibodies cross-react with Hsp60. Therefore, the aim of our present study was to analyze the cross-reactivity of anti-CRP antibodies (Ab) with Hsp60 in solid-phase enzyme immune assays, in epitope studies using a series of overlapping synthetic peptides, and in Ouchterlony analyses. We found that three different commercial rabbit polyclonal antibodies and two monoclonal (9C9 and CRP-8) anti-CRP antibodies specifically recognize recombinant human Hsp60 and recombinantMycobacterium tuberculosisHsp65, respectively. Hsp60 was found to inhibit the binding of anti-CRP polyclonal Ab to Hsp60. Six epitope regions of Hsp60 were recognized by the anti-CRP antibodies, and one region (amino acids [AA] 218 to 232) was recognized by monoclonal antibodies CRP-8 and 9C9. This epitope region of Hsp60 displays 26.6% amino acid identity to CRP AA region 77 to 90. These data suggest that the B-cell epitopes shared between CRP and Hsp60 give rise to a true mimicry-based cross-reaction and the induction of cross-reactive antibodies. Our study underlines the importance of thorough study design and careful interpretation of results while using polyclonal anti-CRP antibodies for histochemistry, especially at low dilutions. Furthermore, analytical interference with Hsp60 in CRP assays should also be tested.
A Human Monoclonal Immunoglobulin M Reduces Bacteremia and Inflammation in a Mouse Model of Systemic Pneumococcal Infection
Authors: Fabrizio Kevin , Groner Abraham , Boes Marianne , Pirofski Liise-anne 
Published: February 2007

ABSTRACTAntibody-based approaches to pneumococcal disease may hold promise for immunocompromised patients in whom vaccines are less immunogenic and/or in the context of antimicrobial resistance. Antibody-mediated protection against experimental pneumococcal pneumonia has been shown to depend on immunoregulation, but the relationship between antibody and protection against pneumococcal sepsis and immunoregulation has not been examined. Similarly, the requirement for B and T cells for antibody efficacy is not known. In this study, we determined the efficacy of the human pneumococcal capsular polysaccharide serotype 3-specific antibody, A7 (immunoglobulin M [IgM]), in secretory IgM (sIgM)−/−, CD4−/−, CD8−/−, μMT−/−, and SCID mice and investigated its effect on cytokine and chemokine expression in sera and spleens from mice with intact cellular immunity. A7 is known to be protective against systemic infection with serotype 3 and to require complement for efficacy. Compared to that of an isotype control antibody, A7 administration prolonged the survival of mice of each immunodeficient strain and was associated with a significant reduction in CFU in blood, lung, and spleen samples and a significantly reduced level of keratinocyte-derived chemokine (KC), interleukin-6 (IL-6), and macrophage inflammatory protein-2 (MIP-2) expression in normal and sIgM−/−mice. Studies with mice treated with penicillin revealed similar reductions in CFU and similar levels of IL-6, KC, or MIP-2 expression in A7- and penicillin-treated mice. These findings demonstrate that natural IgM and B and T cells are dispensable for A7-mediated protection against experimental pneumococcal sepsis and suggest that the efficacy of antibody-mediated protection depends on immunomodulation. Taken together, our data extend the association between antibody-mediated protection and immunomodulation to protection against systemic pneumococcal infection and to a clinically important serotype often responsible for pneumococcal sepsis.
Limited Protection Conferred by a DNA Vaccine against a Lethal Pseudorabies Virus Infection at Day 5 Postvaccination
Authors: Dory Daniel , Torché Anne-Marie , Béven Véronique , Cariolet Roland , Jestin André 
Published: February 2007

ABSTRACTNo pseudorabies virus (PRV)-specific neutralizing or immunoglobulin G1-type antibodies were detected in sera 5 days after injection of a DNA vaccine against PRV infection in pigs. PRV-stimulated peripheral blood mononuclear cells produced gamma interferon mRNA in vitro. Two out of five pigs recovered from lethal PRV infection without attenuation of nasal viral excretion.
In Vivo Expression of and Cell-Mediated Immune Responses to the Plasmid-Encoded Virulence-Associated Proteins ofRhodococcus equiin Foals
Authors: Jacks Stephanie , Giguère Steeve , Prescott John F. 
Published: February 2007

ABSTRACTRhodococcus equiis a facultative intracellular pathogen that causes pneumonia in foals but does not induce disease in adult horses. Virulence ofR. equidepends on the presence of a large plasmid, which encodes a family of seven virulence-associated proteins (VapA and VapC to VapH). Eradication ofR. equifrom the lungs depends on gamma interferon (IFN-γ) production by T lymphocytes. The objectives of the present study were to determine the relative in vivo expression of thevapgenes ofR. equiin the lungs of infected foals, to determine the recall response of bronchial lymph node (BLN) lymphocytes from foals and adult horses to each of the Vap proteins, and to compare the cytokine profiles of proliferating lymphocytes between foals and adult horses.vapA,vapD, andvapGwere preferentially expressed in the lungs of infected foals, and expression of these genes in the lungs was significantly (P< 0.05) higher than that achieved during in vitro growth. VapA and VapC induced the strongest lymphoproliferative responses for foals and adult horses. There was no significant difference in recall lymphoproliferative responses or IFN-γ mRNA expression by bronchial lymph node lymphocytes between foals and adults. In contrast, interleukin 4 (IL-4) expression was significantly higher for adults than for foals for each of the Vap proteins. The ratio of IFN-γ to IL-4 was significantly higher for foals than for adult horses for most Vap proteins. Therefore, foals are immunocompetent and are capable of mounting lymphoproliferative responses of the same magnitude and cytokine phenotype as those of adult horses.
Comparison of Mantoux and QuantiFERON TB Gold Tests for Diagnosis of Latent Tuberculosis Infection in Army Personnel
Authors: Franken Willeke P. J. , Timmermans Joost F. , Prins Corine , Slootman Evert-Jan H. J. , Dreverman Johan , Bruins Hans , van Dissel Jaap T. , Arend Sandra M. 
Published: February 2007

ABSTRACT

The tuberculin skin test (TST) was compared with QuantiFERON-TB Gold in-tube (QFT-GIT) test for the diagnosis of tuberculosis in non-
Mycobacterium bovis
BCG-vaccinated military personnel. Among subjects positive by TST, 44.4% of recruits were positive by QFT-GIT compared with 11.5% subjects tested after missions abroad, suggesting that most TST conversions in the latter group were caused by nontuberculous mycobacteria.
Low Maternal Viral Loads and Reduced Granulocyte-Macrophage Colony-Stimulating Factor Levels Characterize Exposed, Uninfected Infants Who Develop Protective Human Immunodeficiency Virus Type 1-Specific Responses
Authors: Schramm Diana B. , Meddows-Taylor Stephen , Gray Glenda E. , Kuhn Louise , Tiemessen Caroline T. 
Published: February 2007

ABSTRACTHuman immunodeficiency virus type 1 (HIV-1)-specific cellular immune responses are elicited in a proportion of infants born to HIV-1-infected mothers and are associated with protection against vertical transmission. To investigate correlates of these HIV-1-specific responses, we examined levels of the immune activation markers neopterin, β2-microglobulin (β2-m), and solublel-selectin (sl-selectin); the immunomodulatory and hematopoietic factors interleukin-7 (IL-7), stromal-cell-derived factor 1 alpha (CXCL12), and granulocyte-macrophage colony-stimulating factor (GM-CSF); and the immunoregulatory cytokine IL-10 among a group of newborns born to HIV-1-positive mothers who did not receive any antiretroviral drugs for prevention of perinatal HIV-1 transmission. Cellular immune responses to HIV-1 envelope (Env) peptides were also measured. We aimed to determine whether newborns who elicit HIV-1-specific cellular immune responses (Env+) and those who lack these responses (Env−) exhibit unique immune features. Our data confirmed that no Env+infants acquired HIV-1 infection. Among exposed, uninfected infants, Env+infants had reduced immune activation (as measured by β2-m and sl-selectin levels in cord blood plasma) compared to Env−infants as well as reduced GM-CSF levels in cord blood plasma. There was also a reduced ability of cord blood mononuclear cells to be induced to produce GM-CSF among Env+infants. Maternal viral load was lower in Env+infants, suggesting that exposure to low levels of antigen may be responsible for priming the protective responses. These findings suggest that infants who are able to develop apparently protective HIV-1-specific cellular immune responses have immunological features and viral exposure histories that distinguish them from their nonresponder counterparts, providing new insights into the development of HIV-1 protective immunity.
Molecular Analysis of Oral and Respiratory Bacterial Species Associated with Ventilator-Associated Pneumonia
Authors: Bahrani-Mougeot Farah K. , Paster Bruce J. , Coleman Shirley , Barbuto Sara , Brennan Michael T. , Noll Jenene , Kennedy Thomas , Fox Philip C. , Lockhart Peter B. 
Published: February 2007

ABSTRACT

Trauma intensive care unit (TICU) patients requiring mechanical respiratory support frequently develop ventilator-associated pneumonia (VAP). Oral and oropharyngeal bacteria are believed to be responsible for many cases of VAP, but definitive evidence of this relationship is lacking. Earlier studies used conventional culture-based methods for identification of bacterial pathogens, but these methods are insufficient, as some bacteria may be uncultivable or difficult to grow. The purpose of this study was to use a culture-independent molecular approach to analyze and compare the bacterial species colonizing the oral cavity and the lungs of TICU patients who developed VAP. Bacterial samples were acquired from the dorsal tongue and bronchoalveolar lavage fluid of 16 patients. Bacterial DNA was extracted, and the 16S rRNA genes were PCR amplified, cloned into
Escherichia coli
, and sequenced. The sequencing data revealed the following: (i) a wide diversity of bacterial species in both the oral and pulmonary sites, some of them novel; (ii) known and putative respiratory pathogens colonizing both the oral cavity and lungs of 14 patients; and (iii) a number of bacterial pathogens (e.g.,
Dialister pneumosintes
,
Haemophilus segnis
,
Gemella morbillorum
, and
Pseudomonas fluorescens
) in lung samples that had not been reported previously at this site when culture-based methods were used. Our data indicate that the dorsal surface of the tongue serves as a potential reservoir for bacterial species involved in VAP. Furthermore, it is clear that the diversity of bacterial pathogens for VAP is far more complex than the current literature suggests.
Colorimetric Phage-Based Assay for Detection of Rifampin-Resistant
Mycobacterium tuberculosis
Authors: McNerney Ruth , Mallard Kim , Urassa Honorathy M. R. , Lemma Eshetu , Donoghue Helen D. 
Published: February 2007

ABSTRACT

Tests based on bacteriophage replication enable rapid screening of
Mycobacterium tuberculosis
for drug resistance. We describe a novel broth-based colorimetric method for detecting phage replication. When clinical isolates were tested by this novel method, high concordance was observed with both the traditional phage assay and gene mutation analysis for detection of resistance to rifampin.
Evaluation of Etest and Disk Diffusion Methods Compared with Broth Microdilution Antifungal Susceptibility Testing of Clinical Isolates of
Candida
spp. against Posaconazole
Authors: Diekema Daniel J. , Messer Shawn A. , Hollis Richard J. , Boyken Linda B. , Tendolkar Shailesh , Kroeger Jennifer , Pfaller Michael A. 
Published: February 2007

ABSTRACT

We performed Etest, disk diffusion, and broth microdilution susceptibility testing of 2,171 clinical isolates of
Candida
spp. against posaconazole. By using provisional breakpoints for comparison purposes only, the categorical agreement between the agar-based methods and broth microdilution results ranged from 93 to 98%, with <1% very major errors. The essential agreement (within 2 well dilutions) between the Etest and broth microdilution methods was 94%. These agar-based methods hold promise as simple and reliable methods for determination of the posaconzole susceptibilities of
Candida
spp.
Determination of
bft
Gene Subtypes in
Bacteroides fragilis
Clinical Isolates
Authors: Avila-Campos Mario J. , Liu Chengxu , Song Yuli , Rowlinson Marie-Claire , Finegold Sydney M. 
Published: February 2007

ABSTRACT

A rapid multiplex PCR approach was developed to detect the
bft
gene subtypes in
Bacteroides fragilis
clinical isolates. This technique could be used to look at the epidemiology of enterotoxigenic strains of
B. fragilis
in clinical infections and whether there is a correlation between disease and the presence of
B. fragilis
enterotoxin.
Comparison of the MChip to Viral Culture, Reverse Transcription-PCR, and the QuickVue Influenza A+B Test for Rapid Diagnosis of Influenza
Authors: Mehlmann Martin , Bonner Aleta B. , Williams John V. , Dankbar Daniela M. , Moore Chad L. , Kuchta Robert D. , Podsiad Amy B. , Tamerius John D. , Dawson Erica D. , Rowlen Kathy L. 
Published: February 2007

ABSTRACT
The performance of a diagnostic microarray (the MChip assay) for influenza was compared in a blind study to that of viral culture, reverse transcription (RT)-PCR, and the QuickVue Influenza A+B test. The patient sample data set was composed of 102 respiratory secretion specimens collected between 29 December 2005 and 2 February 2006 at Scott & White Hospital and Clinic in Temple, Texas. Samples were collected from a wide range of age groups by using direct collection, nasal/nasopharyngeal swabs, or nasopharyngeal aspiration. Viral culture and the QuickVue assay were performed at the Texas site at the time of collection. Aliquots for each sample, identified only by study numbers, were provided to the University of Colorado and Vanderbilt University teams for blinded analysis. When referenced to viral culture, the MChip exhibited a clinical sensitivity of 98% and a clinical specificity of 98%. When referenced to RT-PCR, the MChip assay exhibited a clinical sensitivity of 92% and a clinical specificity of 98%. While the MChip assay currently requires 7 to 8 h to complete the analysis, a significant advantage of the test for influenza virus-positive samples is simultaneous detection and full subtype identification for the two subtypes currently circulating in humans (A/H3N2 and A/H1N1) and avian (A/H5N1) viruses.
Detection of Extended-Spectrum β-Lactamases by Using Boronic Acid as an AmpC β-Lactamase Inhibitor in Clinical Isolates of
Klebsiella
spp. and
Escherichia coli
Authors: Song Wonkeun , Bae Il Kwon , Lee You-Nae , Lee Chae-Hoon , Lee Sang Hee , Jeong Seok Hoon 
Published: February 2007

ABSTRACT

We evaluated highly sensitive methods using boronic acid (BA) to detect extended-spectrum β-lactamase (ESBL) production. A total of 182 clinical isolates of
Klebsiella
spp. (
n
= 118) and
Escherichia coli
(
n
= 64) were analyzed: 62 harbored only ESBLs, 80 harbored both ESBLs and plasmid-mediated AmpC β-lactamases (pAmpCs), and 40 harbored only pAmpCs. The CLSI confirmatory test detected all isolates that produce only ESBLs but detected 85% of isolates that produce both enzymes. When a ≥5-mm increase in the zone diameter of either the cefotaxime (CTX) or the ceftazidime (CAZ) disk in the presence of both clavulanic acid (CA) and BA was considered to be a positive result, the test detected all isolates that harbor ESBLs (± pAmpCs) but showed frequent false-positive results (50%) for isolates that produce only pAmpCs. Meanwhile, when a ≥3-mm increase in the zone diameter of either the CTX/BA or the CAZ/BA disk in the presence of CA was considered to be a positive result, the test also detected all isolates that harbor ESBLs (± pAmpCs) and showed less frequent false-positive results (5%) in isolates that produce only pAmpCs. The latter new interpretive guideline has enhanced detection of ESBLs in clinical isolates of
Klebsiella
spp. and
Escherichia coli
and allowed detection of an ESBL even when potentially masked by a pAmpC.
First Notification of Positive Blood Cultures and the High Accuracy of the Gram Stain Report
Authors: Søgaard Mette , Nørgaard Mette , Schønheyder Henrik C. 
Published: February 2007

ABSTRACT

When blood cultures turn positive, the attending physicians are usually notified immediately about Gram stain findings. However, information on the accuracy of Gram staining is very limited. We examined the accuracy of preliminary blood culture reports provided by a regional laboratory in an observational study including the years 1996, 2000 to 2001, and 2003. We used data from computer files and technicians' laboratory notes. The study was restricted to cultures with one morphological type. Using cultural identification as a reference, we estimated the sensitivity, specificity, and positive and negative predictive values (PPV and NPV, respectively) for the following defined morphological groups: gram-positive cocci in clusters, gram-positive cocci in chains or diplococci, gram-positive rods, gram-negative cocci, gram-negative rods, and yeasts. We further evaluated the Gram stain and wet mount findings for the most frequent bacterial species/groups. We obtained 5,893 positive blood cultures and the following results for the defined groups: sensitivity, range of 91.3 to 99.7%; specificity, 98.9 to 100%; PPV, 94.6 to 100%; and NPV, 99.0 to 100%. The sensitivity for the most frequent species was in the range 91.3 to 100%, with nonhemolytic streptococci having the lowest value (sensitivity, 91.3%; 95% confidence interval, 86.2 to 94.9%). Wet mount reports were less accurate (sensitivity of 30 to 70% for species with peritrichous motility), and
Enterobacteriaceae
(notably
Salmonella
spp.) accounted for 25% of the reports stating polar motility. In conclusion, we demonstrated a high accuracy of Gram stain reports, whereas wet mount microscopy was generally less accurate.
Role of the Hydrophilic Channels of Simian Virus 40 T-Antigen Helicase in DNA Replication
Authors: Wang Weiping , Manna David , Simmons Daniel T. 
Published: February 2007

ABSTRACT
The simian virus 40 (SV40) hexameric helicase consists of a central channel and six hydrophilic channels located between adjacent large tier domains within each hexamer. To study the function of the hydrophilic channels in SV40 DNA replication, a series of single-point substitutions were introduced at sites not directly involved in protein-protein contacts. The mutants were characterized biochemically in various ways. All mutants oligomerized normally in the absence of DNA. Interestingly, 8 of the 10 mutants failed to unwind an origin-containing DNA fragment and nine of them were totally unable to support SV40 DNA replication in vitro. The mutants fell into four classes based on their biochemical properties. Class A mutants bound DNA normally and had normal ATPase and helicase activities but failed to unwind origin DNA and support SV40 DNA replication. Class B mutants were compromised in single-stranded DNA and origin DNA binding at low protein concentrations. They were defective in helicase activity and unwinding of the origin and in supporting DNA replication. Class C and D mutants possessed higher-than-normal single-stranded DNA binding activity at low protein concentrations. The class C mutants failed to separate origin DNA and support DNA replication. The class D mutants unwound origin DNA normally but were compromised in their ability to support DNA replication. Taken together, these results suggest that the hydrophilic channels have an active role in the unwinding of SV40 DNA from the origin and the placement of the resulting single strands within the helicase.
Participation of Rab5, an Early Endosome Protein, in Hepatitis C Virus RNA Replication Machinery
Authors: Stone Michelle , Jia Shuaizheng , Heo Won Do , Meyer Tobias , Konan Kouacou V. 
Published: February 2007

ABSTRACT
Like most positive-strand RNA viruses, hepatitis C virus (HCV) is believed to replicate its genome on the surface of rearranged membranes. We have shown previously that HCV NS4AB, but not the product NS4B, inhibits endoplasmic reticulum (ER)-to-Golgi protein traffic (K. V. Konan, T. H. Giddings, Jr., M. Ikeda, K. Li, S. M. Lemon, and K. Kirkegaard, J. Virol. 77:7843-7855). However, both NS4AB and NS4B can induce “membranous web” formation, first reported by Egger et al. (D. B Egger, R. Gosert, L. Bianchi, H. E. Blum, D. Moradpour, and K. Bienz, J. Virol. 76:5974-5984), which is also observed in HCV-infected cells (Y. Rouille, F. Helle, D. Delgrange, P. Roingeard, C. Voisset, E. Blanchard, S. Belouzard, J. McKeating, A. H. Patel, G. Maertens, T. Wakita, C. Wychowski, and J. Dubuisson, J. Virol. 80:2832-2841) and cells that bear a subgenomic NS5A-green fluorescent protein (GFP) replicon (D. Moradpour, M. J. Evans, R. Gosert, Z. Yuan, H. E. Blum, S. P. Goff, B. D. Lindenbach, and C. M. Rice, J. Virol. 78:7400-7409). To determine the intracellular origin of the web, we examined NS4B colocalization with endogenous cellular markers in the context of the full-length or subgenomic replicon. We found that, in addition to ER markers, early endosome (EE) proteins, including Rab5, were associated with web-inducing protein NS4B. Furthermore, an immunoisolated fraction containing NS4B was found to contain both ER and EE proteins. Using fluorescence microscopy, we showed that wild-type and constitutively active Rab5 proteins were associated with NS4B. Interestingly, expression of dominant-negative Rab5 resulted in significant loss of GFP fluorescence in NS5A-GFP replicon cells. We also found that a small reduction in Rab5 protein expression decreased HCV RNA synthesis significantly. Furthermore, transfection of labeled Rab5 small interfering RNAs into NS5A-GFP replicon cells resulted in a significant decrease in GFP fluorescence. Finally, Rab5 protein was found to coimmunoprecipitate with HCV NS4B. These studies suggest that EE proteins, including Rab5, may play a role in HCV genome replication or web formation.