bluuebunny/crossref_metadata_2025_split huggingface dataset:
Binding to Gangliosides Containing
N
-Acetylneuraminic Acid Is Sufficient To Mediate the Immunomodulatory Properties of the Nontoxic Mucosal Adjuvant LT-IIb(T13I)
Authors: Nawar Hesham F. , Berenson Charles S. , Hajishengallis George , Takematsu Hiromu , Mandell Lorrie , Clare Ragina L. , Connell Terry D. 
Published: April 2010

ABSTRACT

By use of a mouse mucosal immunization model, LT-IIb(T13I), a nontoxic mutant type II heat-labile enterotoxin, was shown to have potent mucosal and systemic adjuvant properties. In contrast to LT-IIb, which binds strongly to ganglioside receptors decorated with either
N
-acetylneuraminic acid (NeuAc) or
N
-glycolylneuraminic acid (NeuGc), LT-IIb(T13I) binds NeuAc gangliosides much less well. Rather, LT-IIb(T13I) binds preferentially to NeuGc gangliosides. To determine if the adjuvant properties of LT-IIb(T13I) are altered in the absence of NeuGc ganglioside receptors, experiments were conducted using a
Cmah
-null mouse line which is deficient in the synthesis of NeuGc gangliosides. Several immunomodulatory properties of LT-IIb(T13I) were shown to be dependent on NeuGc gangliosides. LT-IIb(T13I) had reduced binding activity for NeuGc-deficient B cells and macrophages; binding to NeuGc-deficient T cells and dendritic cells (DC) was essentially undetectable. Treatment of
Cmah
-null macrophages with LT-IIb(T13I), however, upregulated the transcription of interleukin-4 (IL-4), IL-6, IL-17, and gamma interferon (IFN-γ), four cytokines important for promoting immune responses. The production of mucosal IgA and serum IgG against an immunizing antigen was augmented in NeuGc-deficient mice administered LT-IIb(T13I) as a mucosal adjuvant. Notably, NeuGc gangliosides are not expressed in humans. Still, treatment of human monocytes with LT-IIb(T13I) induced the secretion of IL-6, an inflammatory cytokine that mediates differential control of leukocyte activation. These results suggested that NeuAc gangliosides are sufficient to mediate the immunomodulatory properties of LT-IIb(T13I) in mice and in human cells. The nontoxic mutant enterotoxin LT-IIb(T13I), therefore, is potentially a new and safe human mucosal adjuvant.
Rapid, Simple, Quantitative, and Highly Sensitive Antibody Detection for Lyme Disease
Authors: Burbelo Peter D. , Issa Alexandra T. , Ching Kathryn H. , Cohen Jeffrey I. , Iadarola Michael J. , Marques Adriana 
Published: April 2010

ABSTRACTThere is currently a need for improved serological tests for the diagnosis and monitoring of Lyme disease, an infection caused byBorrelia burgdorferi. In the present study, we evaluated luciferase immunoprecipitation systems (LIPSs) for use for profiling of the antibody responses to a panel ofB. burgdorferiproteins for the diagnosis of Lyme disease. Initially, serum samples from a cohort of patients and controls (n= 46) were used for training and were profiled by the use of 15 differentB. burgdorferiantigen constructs. For the patient sera, the antibody responses to severalB. burgdorferiantigens, including VlsE, flagellin (FlaB), BmpA, DbpA, and DbpB, indicated that the antigens had high levels of immunoreactivity. However, the best diagnostic performance was achieved with a synthetic protein, designated VOVO, consisting of a repeated antigenic peptide sequence, VlsE-OspC-VlsE-OspC, Analysis of an independent set of serum samples (n= 139) used for validation showed that the VOVO LIPS test had 98% sensitivity (95% confidence interval [CI], 93% to 100%;P< 0.0001) and 100% specificity (95% CI, 94% to 100%;P< 0.0001). Similarly, the C6 peptide enzyme-linked immunosorbent assay (ELISA) also had 98% sensitivity (95% CI, 93% to 100%;P< 0.0001) and 98% specificity (95% CI, 90% to 100%;P< 0.0001). Receiver operating characteristic analysis revealed that the rates of detection of Lyme disease by the LIPS test and the C6 ELISA were not statistically different. However, the VOVO LIPS test displayed a wide dynamic range of antibody detection spanning over 10,000-fold without the need for serum dilution. These results suggest that screening by the LIPS test with VOVO and otherB. burgdorferiantigens offers an efficient quantitative approach for evaluation of the antibody responses in patients with Lyme disease.
Molecular Evidence of Perinatal Transmission of
Bartonella vinsonii
subsp.
berkhoffii
and
Bartonella henselae
to a Child
Authors: Breitschwerdt Edward B. , Maggi Ricardo G. , Farmer Peter , Mascarelli Patricia E. 
Published: April 2010

ABSTRACT

Bartonella vinsonii
subsp.
berkhoffii
,
Bartonella henselae
, or DNA of both organisms was amplified and sequenced from blood, enrichment blood cultures, or autopsy tissues from four family members. Historical and microbiological results support perinatal transmission of
Bartonella
species in this family. It is of clinical relevance that
Bartonella
spp. may adversely influence human reproductive performance.
Cardiac Tamponade and Heart Failure Due to Myopericarditis as a Presentation of Infection with the Pandemic H1N1 2009 Influenza A Virus
Authors: Puzelli Simona , Buonaguro Franco M. , Facchini Marzia , Palmieri Annapina , Calzoletti Laura , De Marco Maria A. , Arace Pasquale , de Campora Enrico , Esposito Ciro , Cassone Antonio , Rezza Giovanni , Donatelli Isabella 
Published: April 2010

ABSTRACTWe describe a fatal case of myopericarditis presenting with cardiac tamponade in a previously healthy 11-year-old child. Pandemic H1N1 2009 influenza A virus sequences were identified in throat and myocardial tissues and pericardial fluid, suggesting damage of myocardial cells directly caused by the virus.
Improving Molecular Detection of Fungal DNA in Formalin-Fixed Paraffin-Embedded Tissues: Comparison of Five Tissue DNA Extraction Methods Using Panfungal PCR
Authors: Muñoz-Cadavid C. , Rudd S. , Zaki S. R. , Patel M. , Moser S. A. , Brandt M. E. , Gómez B. L. 
Published: April 2010

ABSTRACT
DNA extraction from formalin-fixed paraffin-embedded (FFPE) tissues is difficult and requires special protocols in order to extract small amounts of DNA suitable for amplification. Most described methods report an amplification success rate between 60 and 80%; therefore, there is a need to improve molecular detection and identification of fungi in FFPE tissue. Eighty-one archived FFPE tissues with a positive Gomori methenamine silver (GMS) stain were evaluated using five different commercial DNA extraction kits with some modifications. Three different panfungal PCR assays were used to detect fungal DNA, and two housekeeping genes were used to assess the presence of amplifiable DNA and to detect PCR inhibitors. The sensitivities of the five extraction protocols were compared, and the quality of DNA detection (calculated for each kit as the number of housekeeping gene PCR-positive samples divided by the total number of samples) was 60 to 91% among the five protocols. The efficiencies of the three different panfungals used (calculated as the number of panfungal-PCR-positive samples divided by the number of housekeeping gene PCR-positive samples) were 58 to 93%. The panfungal PCR using internal transcribed spacer 3 (ITS3) and ITS4 primers yielded a product in most FFPE tissues. Two of the five DNA extraction kits (from TaKaRa and Qiagen) showed similar and promising results. However, one method (TaKaRa) could extract fungal DNA from 69 of the 74 FFPE tissues from which a housekeeping gene could be amplified and was also cost-effective, with a nonlaborious protocol. Factors such as sensitivity, cost, and labor will help guide the selection of the most appropriate method for the needs of each laboratory.
CagA and VacA Polymorphisms Are Associated with Distinct Pathological Features in
Helicobacter
pylori
-Infected Adults with Peptic Ulcer and Non-Peptic Ulcer Disease
Authors: Panayotopoulou Effrosini G. , Sgouras Dionyssios N. , Papadakos Konstantinos S. , Petraki Kalliopi , Breurec Sébastien , Michopoulos Spyros , Mantzaris Gerassimos , Papatheodoridis George , Mentis Andreas , Archimandritis Athanasios 
Published: April 2010

ABSTRACT

Polymorphic variability in
Helicobacter
pylori
factors CagA and VacA contributes to bacterial virulence. The presence of one CagA EPIYA-C site is an independent risk factor for gastroduodenal ulceration (odds ratio [OR], 4.647; 95% confidence interval [CI], 2.037 to 10.602), while the presence of the
vacA
i1 allele is a risk factor for increased activity (OR, 5.310; 95% CI, 2.295 to 12.287) and severity of gastritis (OR, 3.862; 95% CI, 1.728 to 8.632).
Detection of Inducible Clindamycin Resistance in Beta-Hemolytic Streptococci by Using the CLSI Broth Microdilution Test and Erythromycin-Clindamycin Combinations
Authors: Bowling Jason E. , Owens Aaron E. , McElmeel M. Leticia , Fulcher Letitia C. , Herrera Monica L. , Wickes Brian L. , Jorgensen James H. 
Published: April 2010

ABSTRACT
This study assessed an erythromycin-clindamycin (ERY-CC) broth test for inducible CC resistance in beta-hemolytic streptococci. One hundred one isolates of groups A, B, C, F, and G were tested by the CLSI broth microdilution method. Combinations of 1 and 0.25 μg/ml or 0.5 and 0.25 μg/ml of ERY and CC, respectively, detected all inducible isolates.
Alternative Use for Spectra MRSA Chromogenic Agar in Detection of Methicillin-Resistant
Staphylococcus aureus
from Positive Blood Cultures
Authors: Peterson Jess F. , Dionisio Alexander A. , Riebe Katherine M. , Hall Gerri S. , Wilson Deborah A. , Whittier Susan , DiPersio Joseph R. , Ledeboer Nathan A. 
Published: April 2010

ABSTRACT

Spectra MRSA agar (Remel, Lenexa, KS), a novel chromogenic medium originally developed to detect methicillin-resistant
Staphylococcus aureus
(MRSA) from nasal swabs, was evaluated in this multicenter study for the detection of MRSA from positive blood cultures exhibiting Gram-positive cocci upon initial Gram staining.
Typhoid in Kenya Is Associated with a Dominant Multidrug-Resistant
Salmonella
enterica
Serovar Typhi Haplotype That Is Also Widespread in Southeast Asia
Authors: Kariuki Samuel , Revathi Gunturu , Kiiru John , Mengo Doris M. , Mwituria Joyce , Muyodi Jane , Munyalo Agnes , Teo Yik Y. , Holt Kathryn E. , Kingsley Robert A. , Dougan Gordon 
Published: April 2010

ABSTRACT

In sub-Saharan Africa, the burden of typhoid fever, caused by
Salmonella enterica
serovar Typhi, remains largely unknown, in part because of a lack of blood or bone marrow culture facilities. We characterized a total of 323
S
. Typhi isolates from outbreaks in Kenya over the period 1988 to 2008 for antimicrobial susceptibilities and phylogenetic relationships using single-nucleotide polymorphism (SNP) analysis. There was a dramatic increase in the number and percentage of multidrug-resistant (MDR)
S
. Typhi isolates over the study period. Overall, only 54 (16.7%)
S
. Typhi isolates were fully sensitive, while the majority, 195 (60.4%), were multiply resistant to most commonly available drugs—ampicillin, chloramphenicol, tetracycline, and cotrimoxazole; 74 (22.9%) isolates were resistant to a single antimicrobial, usually ampicillin, cotrimoxazole, or tetracycline. Resistance to these antibiotics was encoded on self-transferrable IncHI1 plasmids of the ST6 sequence type. Of the 94 representative
S
. Typhi isolates selected for genome-wide haplotype analysis, sensitive isolates fell into several phylogenetically different groups, whereas MDR isolates all belonged to a single haplotype, H58, associated with MDR and decreased ciprofloxacin susceptibility, which is also dominant in many parts of Southeast Asia. Derivatives of the same
S
. Typhi lineage, H58, are responsible for multidrug resistance in Kenya and parts of Southeast Asia, suggesting intercontinental spread of a single MDR clone. Given the emergence of this aggressive MDR haplotype, careful selection and monitoring of antibiotic usage will be required in Kenya, and potentially other regions of sub-Saharan Africa.
Evaluation of Bio-Rad MRSA
Select
Agar for Detection of Methicillin-Resistant
Staphylococcus aureus
Directly from Blood Cultures
Authors: Riedel Stefan , Dam Lisa , Stamper Paul D. , Shah Syed A. R. , Carroll Karen C. 
Published: April 2010

ABSTRACT

MRSA
Select
agar (Bio-Rad, Redmond, WA) was evaluated for its performance in detecting MRSA directly from positive blood cultures containing Gram-positive cocci in clusters. Agar plates were evaluated for the presence of pink colonies at 18 to 24 h. Results were compared to organism identification by using standard laboratory methods. Confirming coagulase on pink isolates, the sensitivity and specificity were both 99%.
Direct Identification of Urinary Tract Pathogens from Urine Samples by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry
Authors: Ferreira Laura , Sánchez-Juanes Fernando , González-Ávila Magdalena , Cembrero-Fuciños David , Herrero-Hernández Ana , González-Buitrago José Manuel , Muñoz-Bellido Juan Luis 
Published: April 2010

ABSTRACT

Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been suggested as a reliable method for bacterial identification from cultures. Direct analysis of clinical samples might increase the usefulness of this method, shortening the time for microorganism identification. We compared conventional methods for the diagnosis of urinary tract infections (UTIs) and identification of the urinary tract pathogens (automated screening, plate cultures, and identification based on biochemical characteristics) and a fast method based on conventional screening and MALDI-TOF MS. For this latter method, 4 ml of urine was centrifuged at a low-revolution setting (2,000 ×
g
) to remove leukocytes and then at high revolutions (15,500 ×
g
) to collect bacteria. The pellet was washed and then applied directly to the MALDI-TOF MS plate. Two hundred sixty urine samples, detected as positive by the screening device (UF-1000i), were processed by culture and MALDI-TOF MS. Twenty samples were positive in the screening device but negative in culture, and all of them were also negative by MALDI-TOF MS. Two-hundred thirty-five samples displayed significant growth of a single morphological type in culture. Two-hundred twenty of them showed bacterial growth of >10
5
CFU/ml. Microorganism identifications in this group were coincident at the species level in 202 cases (91.8%) and at the genus level in 204 cases (92.7%). The most frequent microorganism was
Escherichia coli
(173 isolates). MALDI-TOF MS identified this microorganism directly from the urine sample in 163 cases (94.2%). Our results show that MALDI-TOF MS allows bacterial identification directly from infected urine in a short time, with high accuracy, and especially when Gram-negative bacteria with high bacterial counts are involved.
Molecular Identification of Veterinary Yeast Isolates by Use of Sequence-Based Analysis of the D1/D2 Region of the Large Ribosomal Subunit
Authors: Garner Cherilyn D. , Starr Jennifer K. , McDonough Patrick L. , Altier Craig 
Published: April 2010

ABSTRACT

Conventional methods of yeast identification are often time-consuming and difficult; however, recent studies of sequence-based identification methods have shown promise. Additionally, little is known about the diversity of yeasts identified from various animal species in veterinary diagnostic laboratories. Therefore, in this study, we examined three methods of identification by using 109 yeast samples isolated during a 1-year period from veterinary clinical samples. Comparison of the three methods—traditional substrate assimilation, fatty acid profile analysis, and sequence-based analysis of the region spanning the D1 and D2 regions (D1/D2) of the large ribosomal subunit—showed that sequence analysis provided the highest percent identification among the three. Sequence analysis identified 87% of isolates to the species level, whereas substrate assimilation and fatty acid profile analysis identified only 54% and 47%, respectively. Less-stringent criteria for identification increased the percentage of isolates identified to 98% for sequence analysis, 62% for substrate assimilation, and 55% for fatty acid profile analysis. We also found that sequence analysis of the internal transcribed spacer 2 (ITS2) region provided further identification for 36% of yeast not identified to the species level by D1/D2 sequence analysis. Additionally, we identified a large variety of yeast from animal sources, with at least 30 different species among the isolates tested, and with the majority not belonging to the common
Candida
spp., such as
C. albicans
,
C. glabrata
,
C. tropicalis
, and the
C. parapsilosis
group. Thus, we determined that sequence analysis of the D1/D2 region was the best method for identification of the variety of yeasts found in a veterinary population.
Development and Evaluation of anompAQuantitative Real-Time PCR Assay forChlamydia trachomatisSerovar Determination
Authors: Stevens Matthew P. , Twin Jimmy , Fairley Christopher K. , Donovan Basil , Tan Sarah E. , Yu Jingxi , Garland Suzanne M. , Tabrizi Sepehr N. 
Published: April 2010

ABSTRACTKnowledge of circulatingChlamydia trachomatisserovars can be beneficial for sexual network surveillance, monitoring treatment success, and associating specific clinical manifestations. Typically,C. trachomatisserovars are predicted by nucleotide sequencing of four variable domains within theompAgene. However, sequencing procedures can be labor-intensive, are not readily available, and can lack the capacity to identify multiple serovars. This study describes the development and evaluation of a quantitative real-time PCR (qPCR) test algorithm for the rapid prediction ofC. trachomatisserovars, including ocular (A to C) and anogenital (D to L3) strains. This test comprises a primary qPCR to confirmC. trachomatispositivity and the phylogenetic group(s) present and a secondary set of qPCRs to determine specific serovars. Cell culture isolates from 15 prototypicC. trachomatisserovars were correctly identified using this assay, with no cross-reactivity observed among serovars or with other common pathogenic microorganisms. Five hundred clinical specimens (previously diagnosed as beingC. trachomatispositive) were evaluated by qPCR, with their results compared to results obtained by conventional sequencing. The qPCR identified 88.9% (423/476) complete matches (95% confidence interval [CI], 86 to 92%) of serovars compared to the results obtained using the sequence-based approach. Among the anogenital specimens, 2.4% (12/494) (95% CI, 1.3 to 4.2%) contained multiple serovars, categorized as single-serovar infections by conventional sequencing. Overall, this test exhibited high discriminatory success for predictingC. trachomatisserovars, particularly among anogenital infections. This is the first report of a qPCR typing assay offering differentiation ofC. trachomatisserovars associated with both anogenital and ocular diseases.
Rapid Antifungal Susceptibility Determination for Yeast Isolates by Use of Etest Performed Directly on Blood Samples from Patients with Fungemia
Authors: Guinea Jesús , Recio Sandra , Escribano Pilar , Torres-Narbona Marta , Peláez Teresa , Sánchez-Carrillo Carlos , Rodríguez-Créixems Marta , Bouza Emilio 
Published: April 2010

ABSTRACT

We prospectively determined the antifungal susceptibility of yeast isolates causing fungemia using the Etest on direct blood samples (195 prospectively collected and 133 laboratory prepared). We compared the Etest direct (24 h of incubation) with CLSI M27-A3 and the standard Etest methodologies for fluconazole, voriconazole, posaconazole, isavuconazole, caspofungin, and amphotericin B. Strains were classified as susceptible, resistant, or nonsusceptible using CLSI breakpoints (voriconazole breakpoints were used for posaconazole and isavuconazole). Categorical errors between Etest direct and CLSI M27-A3 for azoles were mostly minor. No errors were detected for caspofungin, and high percentages of major errors were detected for amphotericin B. For the azoles, false susceptibility (very major errors) was found in only two (0.6%) isolates (
Candida tropicalis
and
C. glabrata
). False resistance (major errors) was detected in 46 (14%) isolates for the three azoles (in 23 [7%] after excluding posaconazole). Etest direct of posaconazole yielded a higher number of major errors than the remaining azoles, especially for
C. glabrata
,
Candida
spp., and other yeasts. Excluding
C. glabrata
,
Candida
spp., and other yeasts, the remaining species did not yield major errors. Etest direct for fluconazole, voriconazole, isavuconazole, and caspofungin shows potential as an alternative to the CLSI M27-A3 procedure for performing rapid antifungal susceptibility tests on yeast isolates from patients with fungemia. Etest direct is a useful tool to screen for the presence of azole-resistant and caspofungin-nonsusceptible strains.
Evaluation of a New Selective Medium, BD BBL CHROMagar MRSA II, for Detection of Methicillin-Resistant
Staphylococcus aureus
in Different Specimens
Authors: Wendt C. , Havill N. L. , Chapin K. C. , Boyce J. M. , Dickenson R. , Eigner U. , Schütt S. , Fahr A. M. 
Published: April 2010

ABSTRACT

The sensitivity of screening for methicillin-resistant
Staphylococcus aureus
(MRSA) can be improved by adding other specimen sites to nares. We describe an evaluation of a new selective medium, BBL CHROMagar MRSA II (CMRSAII), for its ability to detect MRSA from different specimen types. CMRSAII is a chromogenic medium which incorporates cefoxitin for the detection of MRSA. A study was performed at four clinical laboratories with the following specimens: 1,446 respiratory, 694 stool, 1,275 skin, and 948 wound specimens and 688 blood culture bottles containing Gram-positive cocci. The recovery of MRSA on traditional culture media was compared to results with CMRSAII.
S. aureus
was tested by cefoxitin disk diffusion. CMRSAII was interpreted as positive for MRSA at 24 h (range, 18 to 28 h) based solely on the visualization of mauve-colored colonies and at 48 h (range, 36 to 52 h) based on detection of mauve colonies with subsequent confirmation as
S. aureus
(by coagulase or latex agglutination testing). MRSA was recovered more frequently on CMRSAII (89.8% at 24 h and 95.6% at 48 h) than on traditional culture plates (83.1% at 24 h and 79.8% at 48 h) for all specimen types combined (
P
< 0.001). The percent sensitivities of CMRSAII at 24- and 48-h reads, respectively, were 85.5 and 92.4% for respiratory specimens, 87.9% and 98.3% for stool specimens, 88.4% and 96.1% for skin specimens, 92.1% and 94.6% for wound specimens, and 100% and 100% for positive blood cultures. The specificity was 99.8% for respiratory specimens and 100% for all others. In conclusion, CMRSAII is a reliable screening medium for multiple specimen types.
Evaluation of a New Selective Medium, BD BBL CHROMagar MRSA II, for Detection of Methicillin-Resistant
Staphylococcus aureus
in Stool Specimens
Authors: Havill Nancy L. , Boyce John M. 
Published: April 2010

ABSTRACT

We compared the recovery of methicillin-resistant
Staphylococcus aureus
(MRSA) on a new selective chromogenic agar, BD BBL CHROMagar MRSA II (CMRSAII), to that on traditional culture media with 293 stool specimens. The recovery of MRSA was greater on the CMRSAII agar. Screening of stool samples can identify patients who were previously unknown carriers of MRSA.