bluuebunny/crossref_metadata_2025_split huggingface dataset:
Evaluation of Three Commercially Available Influenza A Type-Specific Blocking Enzyme-Linked Immunosorbent Assays for Seroepidemiological Studies of Influenza A Virus Infection in Pigs
Authors: Tse Maying , Kim Mia , Chan Chung-Hei , Ho Po-Lai , Ma Siu-Kit , Guan Yi , Peiris J. S. M. 
Published: January 2012

ABSTRACTThe reverse zoonotic transmission of the pandemic H1N1 2009 influenza virus to swine necessitates enhanced surveillance of swine for influenza virus infection. Using a well-characterized panel of naturally infected swine sera, we evaluated and optimized the performances of three commercially available competitive enzyme-linked immunosorbent assays (ELISAs), namely, the IDEXX Influenza A Ab test, IDEXX AI MultiS-Screen Ab test, and IDVet ID Screen influenza A antibody competition ELISA, for detecting influenza A virus-reactive antibodies in swine. Receiver operating characteristic (ROC) analysis suggests that adjustment of the manufacturer-recommended cutoff values optimizes the sensitivity and specificity of these assays, making them applicable for seroepidemiology studies of swine influenza. Using such optimized cutoff levels, the sensitivity and specificity of the IDEXX Influenza A Ab test were 86% and 89%, respectively; those for the IDEXX AI MultiS-Screen Ab test were 91% and 87%, respectively; and those for the IDVet ID Screen influenza A test were 95% and 79%, respectively.
Molecular Characterization of Toxoplasma gondii Formin 3, an Actin Nucleator Dispensable for Tachyzoite Growth and Motility
Authors: Daher Wassim , Klages Natacha , Carlier Marie-France , Soldati-Favre Dominique 
Published: December 2011

ABSTRACT

Toxoplasma gondii
belongs to the phylum Apicomplexa, a group of obligate intracellular parasites that rely on gliding motility to enter host cells. Drugs interfering with the actin cytoskeleton block parasite motility, host cell invasion, and egress from infected cells. Myosin A, profilin, formin 1, formin 2, and actin-depolymerizing factor have all been implicated in parasite motility, yet little is known regarding the importance of actin polymerization and other myosins for the remaining steps of the parasite lytic cycle. Here we establish that
T. gondii
formin 3 (TgFRM3), a newly described formin homology 2 domain (FH2)-containing protein, binds to
Toxoplasma
actin and nucleates rabbit actin assembly
in vitro
. TgFRM3 expressed as a transgene exhibits a patchy localization at several distinct structures within the parasite. Disruption of the
TgFRM3
gene by double homologous recombination in a
ku80-ko
strain reveals no vital function for tachyzoite propagation
in vitro
, which is consistent with its weak level of expression in this life stage. Conditional stabilization of truncated forms of TgFRM3 suggests that different regions of the molecule contribute to distinct localizations. Moreover, expression of TgFRM3 lacking the C-terminal domain severely affects parasite growth and replication. This work provides a first insight into how this specialized formin, restricted to the group of coccidia, completes its actin-nucleating activity.
Rapamycin Exerts Antifungal Activity
In Vitro
and
In Vivo
against Mucor circinelloides via FKBP12-Dependent Inhibition of Tor
Authors: Bastidas Robert J. , Shertz Cecelia A. , Lee Soo Chan , Heitman Joseph , Cardenas Maria E. 
Published: December 2011

ABSTRACT

The zygomycete
Mucor circinelloides
is an opportunistic fungal pathogen that commonly infects patients with malignancies, diabetes mellitus, and solid organ transplants. Despite the widespread use of antifungal therapy in the management of zygomycosis, the incidence of infections continues to rise among immunocompromised individuals. In this study, we established that the target and mechanism of antifungal action of the immunosuppressant rapamycin in
M. circinelloides
are mediated via conserved complexes with FKBP12 and a Tor homolog. We found that spontaneous mutations that disrupted conserved residues in FKBP12 conferred rapamycin and FK506 resistance. Disruption of the FKBP12-encoding gene,
fkbA
, also conferred rapamycin and FK506 resistance. Expression of
M. circinelloides
FKBP12 (
Mc
FKBP12) complemented a
Saccharomyces cerevisiae
mutant strain lacking FKBP12 to restore rapamycin sensitivity. Expression of the
Mc
Tor FKBP12-rapamycin binding (FRB) domain conferred rapamycin resistance in
S. cerevisiae
, and
Mc
FKBP12 interacted in a rapamycin-dependent fashion with the
Mc
Tor FRB domain in a yeast two-hybrid assay, validating
Mc
FKBP12 and
Mc
Tor as conserved targets of rapamycin. We showed that
in vitro
, rapamycin exhibited potent growth inhibitory activity against
M. circinelloides
. In a
Galleria mellonella
model of systemic mucormycosis, rapamycin improved survival by 50%, suggesting that rapamycin and nonimmunosuppressive analogs have the potential to be developed as novel antifungal therapies for treatment of patients with mucormycosis.
A 54-Kilodalton Protein Encoded by pBtoxis Is Required for Parasporal Body Structural Integrity in Bacillus thuringiensis subsp. israelensis
Authors: Diaz-Mendoza Mercedes , Bideshi Dennis K. , Federici Brian A. 
Published: December 2011

ABSTRACT

Strains of
Bacillus thuringiensis
such as
B. thuringiensis
subsp.
israelensis
(ONR-60A) and
B. thuringiensis
subsp.
morrisoni
(PG-14) pathogenic for mosquito larvae produce a complex parasporal body consisting of several protein endotoxins synthesized during sporulation that form an aggregate of crystalline inclusions bound together by a multilamellar fibrous matrix. Most studies of these strains focus on the molecular biology of the endotoxins, and although it is known that parasporal body structural integrity is important to achieving high toxicity, virtually nothing is known about the matrix that binds the toxin inclusions together. In the present study, we undertook a proteomic analysis of this matrix to identify proteins that potentially mediate assembly and stability of the parasporal body. In addition to fragments of their known major toxins, namely, Cry4Aa, Cry4Ba, Cry11Aa, and Cyt1Aa, we identified peptides with 100% identity to regions of Bt152, a protein coded for by pBtoxis of
B. thuringiensis
subsp.
israelensis
, the plasmid that encodes all endotoxins of this subspecies. As it is known that the Bt152 gene is expressed in
B. thuringiensis
subsp.
israelensis
, we disrupted its function and showed that inactivation destabilized the parasporal body matrix and, concomitantly, inclusion aggregation. Using fluorescence microscopy, we further demonstrate that Bt152 localizes to the parasporal body in both strains, is absent in other structural or soluble components of the cell, including the endospore and cytoplasm, and in ligand blots binds to purified multilamellar fibrous matrix. Together, the data show that Bt152 is essential for stability of the parasporal body of these strains.
Analysis of the Pseudomonas aeruginosa Regulon Controlled by the Sensor Kinase KinB and Sigma Factor RpoN
Authors: Damron F. Heath , Owings Joshua P. , Okkotsu Yuta , Varga John J. , Schurr Jill R. , Goldberg Joanna B. , Schurr Michael J. , Yu Hongwei D. 
Published: December 2011

ABSTRACT

Alginate overproduction by
Pseudomonas aeruginosa
, also known as mucoidy, is associated with chronic endobronchial infections in cystic fibrosis. Alginate biosynthesis is initiated by the extracytoplasmic function sigma factor (σ
22
; AlgU/AlgT). In the wild-type (wt) nonmucoid strains, such as PAO1, AlgU is sequestered to the cytoplasmic membrane by the anti-sigma factor MucA that inhibits alginate production. One mechanism underlying the conversion to mucoidy is mutation of
mucA
. However, the mucoid conversion can occur in wt
mucA
strains via the degradation of MucA by activated intramembrane proteases AlgW and/or MucP. Previously, we reported that the deletion of the sensor kinase KinB in PAO1 induces an AlgW-dependent proteolysis of MucA, resulting in alginate overproduction. This type of mucoid induction requires the alternate sigma factor RpoN (σ
54
). To determine the RpoN-dependent KinB regulon, microarray and proteomic analyses were performed on a mucoid
kinB
mutant and an isogenic nonmucoid
kinB rpoN
double mutant. In the
kinB
mutant of PAO1, RpoN controlled the expression of approximately 20% of the genome. In addition to alginate biosynthetic and regulatory genes, KinB and RpoN also control a large number of genes including those involved in carbohydrate metabolism, quorum sensing, iron regulation, rhamnolipid production, and motility. In an acute pneumonia murine infection model, BALB/c mice exhibited increased survival when challenged with the
kinB
mutant relative to survival with PAO1 challenge. Together, these data strongly suggest that KinB regulates virulence factors important for the development of acute pneumonia and conversion to mucoidy.
Characterization of Recombinant UDP- and ADP-Glucose Pyrophosphorylases and Glycogen Synthase To Elucidate Glucose-1-Phosphate Partitioning into Oligo- and Polysaccharides in Streptomyces coelicolor
Authors: Asención Diez Matías D. , Peirú Salvador , Demonte Ana M. , Gramajo Hugo , Iglesias Alberto A. 
Published: December 2011

ABSTRACT

Streptomyces coelicolor
exhibits a major secondary metabolism, deriving important amounts of glucose to synthesize pigmented antibiotics. Understanding the pathways occurring in the bacterium with respect to synthesis of oligo- and polysaccharides is of relevance to determine a plausible scenario for the partitioning of glucose-1-phosphate into different metabolic fates. We report the molecular cloning of the genes coding for UDP- and ADP-glucose pyrophosphorylases as well as for glycogen synthase from genomic DNA of
S. coelicolor
A3(2). Each gene was heterologously expressed in
Escherichia coli
cells to produce and purify to electrophoretic homogeneity the respective enzymes. UDP-glucose pyrophosphorylase (UDP-Glc PPase) was characterized as a dimer exhibiting a relatively high
V
max
in catalyzing UDP-glucose synthesis (270 units/mg) and with respect to dTDP-glucose (94 units/mg). ADP-glucose pyrophosphorylase (ADP-Glc PPase) was found to be tetrameric in structure and specific in utilizing ATP as a substrate, reaching similar activities in the directions of ADP-glucose synthesis or pyrophosphorolysis (
V
max
of 0.15 and 0.27 units/mg, respectively). Glycogen synthase was arranged as a dimer and exhibited specificity in the use of ADP-glucose to elongate α-1,4-glucan chains in the polysaccharide. ADP-Glc PPase was the only of the three enzymes exhibiting sensitivity to allosteric regulation by different metabolites. Mannose-6-phosphate, phosphoenolpyruvate, fructose-6-phosphate, and glucose-6-phosphate behaved as major activators, whereas NADPH was a main inhibitor of ADP-Glc PPase. The results support a metabolic picture where glycogen synthesis occurs via ADP-glucose in
S. coelicolor
, with the pathway being strictly regulated in connection with other routes involved with oligo- and polysaccharides, as well as with antibiotic synthesis in the bacterium.
Real-Time PCR-Based Mismatch Amplification Mutation Assay for Specific Detection of CS6-Expressing Allelic Variants of Enterotoxigenic Escherichia coli and Its Application in Assessing Diarrheal Cases and Asymptomatic Controls
Authors: Sabui Subrata , Dutta Sanjucta , Debnath Anusuya , Ghosh Avishek , Hamabata T. , Rajendran K. , Ramamurthy T. , Nataro James P. , Sur Dipika , Levine Myron M. , Chatterjee Nabendu Sekhar 
Published: January 2012

ABSTRACT

Enterotoxigenic
Escherichia coli
(ETEC) expressing the colonization factor CS6 is widespread in many developing countries, including India. The different allelic variants of CS6, caused by point mutations in its structural genes,
cssA
and
cssB
, are designated AIBI, AIIBII, AIIIBI, AIBII, and AIIIBII. A simple, reliable, and specific
m
ismatch
a
mplification
m
utation
a
ssay based on real-time quantitative PCR (MAMA-qPCR) was developed for the first time for the detection of CS6-expressing ETEC, along with the identification of allelic variations. The assay was based on mismatched nucleotide incorporation at the penultimate base at the 3′ ends of the reverse primers specific for
cssA
and
cssB
and was validated using 38 CS6-expressing ETEC isolates. This strategy was effective in detecting all the alleles containing single-nucleotide polymorphisms. Using MAMA-qPCR, we also tested CS6 allelic variants in 145 ETEC isolates from children with acute diarrhea and asymptomatic infections, with the latter serving as controls. We observed that the AIBI and AIIIBI allelic variants were mostly associated with cases rather than controls, whereas the AIIBII variants were detected mostly in controls. In addition, the AIBI and AIIIBI alleles were frequently associated with ETEC harboring the heat-stable toxin gene (
est
) alone or with the heat-labile toxin gene (
elt
), whereas the AIIBII allele was predominant in ETEC isolates harboring the
elt
gene. This study may help in understanding the association of allelic variants in CS6-expressing ETEC with the clinical features of diarrhea, as well as in ETEC vaccine studies.
An Alternative Method for Cultivation of Lawsonia intracellularis
Authors: Vannucci Fabio A. , Wattanaphansak Suphot , Gebhart Connie J. 
Published: January 2012

ABSTRACT

An alternative method for the cultivation of
Lawsonia intracellularis
, an obligate intracellular bacterium and the causative agent of proliferative enteropathy, was developed using an Original Space Bag inflated with a mixture of gas containing 10% hydrogen, 10% carbon dioxide, and 80% nitrogen. The flexibility of this protocol allows the testing of various environmental conditions for static cultivation of this bacterium and the development of diagnostic techniques.
Integrated Analysis of Multiple Gene Expression Profiling Datasets Revealed Novel Gene Signatures and Molecular Markers in Nasopharyngeal Carcinoma
Authors: Huang Chen , Tang Hailin , Zhang Wenling , She Xiaoling , Liao Qianjin , Li Xiaoling , Wu Minghua , Li Guiyuan 
Published: January 2012

AbstractPurpose: To identify the novel gene signatures and molecular markers of nasopharyngeal carcinoma (NPC) by integrated bioinformatics analysis of multiple gene expression profiling datasets.Experimental Design: Seven published gene expression profiling studies and one of our unpublished works were reanalyzed to identify the common significantly dysregulated (CSD) genes in NPC. Overrepresentation analysis of cytogenetic bands, Gene Ontology (GO) categories, pathways were used to explore CSD genes functionally associated with carcinogenesis. The protein expressions of selected CSD genes were examined by immunohistochemistry on tissue microarrays, and the correlations of their expressions with clinical outcomes were evaluated.Results: Using the criteria (genes reported deregulated in more than one study), a total of 962 genes were identified as the CSD genes in NPC. Four upregulated (BUB1B, CCND2, CENPF, and MAD2L1) and two downregulated (LTF and SLPI) genes were markedly reported in six studies. The enrichments of chromosome aberrations were 2q23, 2q31, 7p15, 12q15, 12q22, 18q11, and 18q12 in upregulated genes and 14q32 and 16q13 in downregulated genes. The activated GO categories and pathways related to proliferation, adhesion, invasion, and downregulated immune response had been functionally associated with NPC. SLPI significantly downregulated in nasopharyngeal adenocarcinoma. Furthermore, the high expression of BUB1B or CENPF was associated with poor overall survival of patients.Conclusion: It was first clearly identified the dysregulated expression of BUB1B and SLPI in NPC tissues.Impact: Further studies of the CSD genes as gene signatures and molecular markers of NPC might improve the understanding of the disease and identify new therapeutic targets. Cancer Epidemiol Biomarkers Prev; 21(1); 166–75. ©2011 AACR.
Plasma 25-Hydroxyvitamin D and Risk of Pancreatic Cancer
Authors: Wolpin Brian M. , Ng Kimmie , Bao Ying , Kraft Peter , Stampfer Meir J. , Michaud Dominique S. , Ma Jing , Buring Julie E. , Sesso Howard D. , Lee I-Min , Rifai Nader , Cochrane Barbara B. , Wactawski-Wende Jean , Chlebowski Rowan T. , Willett Walter C. , Manson JoAnn E. , Giovannucci Edward L. , Fuchs Charles S. 
Published: January 2012

Abstract
Background: Laboratory studies suggest that vitamin D may inhibit pancreatic cancer cell growth. However, epidemiologic studies of vitamin D and pancreatic cancer risk have been conflicting.
Methods: To determine whether prediagnostic levels of plasma 25-hydroxyvitamin D (25[OH]D; IDS Inc.; enzyme immunoassay) were associated with risk of pancreatic cancer, we conducted a pooled analysis of nested case–control studies with 451 cases and 1,167 controls from five cohorts through 2008. Median follow-up among controls was 14.1 years in Health Professionals Follow-Up Study (HPFS), 18.3 years in Nurses' Health Study (NHS), 25.3 years in Physicians' Health Study (PHS), 12.2 years in Women's Health Initiative-Observational Study (WHI), and 14.4 years in Women's Health Study (WHS). Logistic regression was used to compare the odds of pancreatic cancer by plasma level of 25(OH)D.
Results: Mean plasma 25(OH)D was lower in cases versus controls (61.3 vs. 64.5 nmol/L, P = 0.005). In logistic regression models, plasma 25(OH)D was inversely associated with odds of pancreatic cancer. Participants in quintiles two through five had multivariable-adjusted ORs (95% confidence intervals) of 0.79 (0.56–1.10), 0.75 (0.53–1.06), 0.68 (0.48–0.97), and 0.67 (0.46–0.97; Ptrend = 0.03), respectively, compared with the bottom quintile. Compared with those with insufficient levels [25[OH]D, &lt;50 nmol/L], ORs were 0.75 (0.58–0.98) for subjects with relative insufficiency [25[OH]D, 50 to &lt;75 nmol/L] and 0.71 (0.52–0.97) for those with sufficient levels [25[OH]D, ≥75 nmol/L]. No increased risk was noted in subjects with 25(OH)D ≥100 nmol/L, as suggested in a prior study. In subgroup analyses, ORs for the top versus bottom quartile of 25(OH)D were 0.72 (0.48–1.08) for women, 0.73 (0.40–1.31) for men, and 0.73 (0.51–1.03) for Whites.
Conclusions: Among participants in five large prospective cohorts, higher plasma levels of 25(OH)D were associated with a lower risk for pancreatic cancer.
Impact: Low circulating 25(OH)D may predispose individuals to the development of pancreatic cancer. Cancer Epidemiol Biomarkers Prev; 21(1); 82–91. ©2011 AACR.
VBM–DTI Correlates of Verbal Intelligence: A Potential Link to Broca's Area
Authors: Konrad Andreas , Vucurevic Goran , Musso Francesco , Winterer Georg 
Published: January 2012

Abstract
Human brain lesion studies first investigated the biological roots of cognitive functions including language in the late 1800s. Neuroimaging studies have reported correlation findings with general intelligence predominantly in fronto-parietal cortical areas. However, there is still little evidence about the relationship between verbal intelligence and structural properties of the brain. We predicted that verbal performance is related to language regions of Broca's and Wernicke's areas. Verbal intelligence quotient (vIQ) was assessed in 30 healthy young subjects. T1-weighted MRI and diffusion tensor imaging data sets were acquired. Voxel-wise regression analyses were used to correlate fractional anisotropy (FA) and mean diffusivity values with vIQ. Moreover, regression analyses of regional brain volume with vIQ were performed adopting voxel-based morphometry (VBM) and ROI methodology. Our analyses revealed a significant negative correlation between vIQ and FA and a significant positive correlation between vIQ and mean diffusivity in the left-hemispheric Broca's area. VBM regression analyses did not show significant results, whereas a subsequent ROI analysis of Broca's area FA peak cluster demonstrated a positive correlation of gray matter volume and vIQ. These findings suggest that cortical thickness in Broca's area contributes to verbal intelligence. Diffusion parameters predicted gray matter ratio in Broca's area more sensitive than VBM methodology.
Redescription and variation of Hyophryne histrio Carvalho, 1954, an enigmatic microhylid frog from the Atlantic Rainforest of Brazil
Authors: Targino Mariane , Pombal José P. 
Published: January 2012

Abstract
Hyophryne histrio was described in 1954 as a new species and placed in its own genus based on a single juvenile specimen from Ilhéus, State of Bahia, northeastern Brazil. This species was known only from the holotype until 1999 when a series of specimens was collected 50 km from the type-locality. Herein, we provide a redescription of the species based on this new material and present new information on morphological variation in order to help with the recognition of this poorly known species. The diagnostic characters are: occipital skin fold present, interdigital membranes between the toes well developed, dark venter with white blotches, an arrow-shaped dorsal color pattern that begins just after the occipital fold, and a distinct pale line between the venter and dorsum. Dermal spines, which were found in all males and vary in the number of spines and location, are reported for the first time in this species; gravid females have dermal spines only in the pericloacal region. We also compare Hyophryne histrio to all other Neotropical microhylid genera.
What’s in a Name? Challenges to the Creation of EU Delegations
Authors: Drieskens Edith 
Published: January 2012

Summary
One of the Lisbon Treaty’s most significant innovations was the creation of the European External Action Service (EEAS), which changed the EU’s functioning not only in Brussels, but also around the world. Zooming in on the multilateral context of the UN in New York, this article examines the new EU delegations and highlights the main challenges that are inherent in their establishment. These delegations could be engrafted upon a wide network of European Commission delegations, yet the literature gives little indication of success in integrating the functions and actors. Adding to the literature and building upon interviews with policy officials in both Brussels and New York, this article indicates an additional external challenge in implementing Lisbon’s provisions, with the context of the UN General Assembly raising more fundamental questions on status and membership — questions that have also shaped the role of the EU delegation to the UN during its first year of operation.
A media comparison study on first aid instruction
Authors: Celik Serkan 
Published: January 2012

Aims: A basic knowledge and understanding of first aid can be invaluable for both teachers and students to be able to provide emergency care in the event of an accident, possibly saving lives and minimizing injury in school settings. The purpose of this research was to perform a comparison of online and face-to-face delivery modes for an introductory first aid course to determine if there existed a difference in student teachers’ success depending on the delivery modes. Methods: The research compares participants’ achievements in a first aid class taught by the same instructor using either face-to-face or online delivery modes. In total, five sections of the course were assigned to either a face-to face or an online treatment condition. Results: The results of a paired samples t-test indicated that student teachers both in experiment and control groups performed adequately to attain the course goals ( p < .05). The difference between the mean scores of all groups in pre- and post-tests verified the achievement of the participant learners. The results of a further independent t-test on groups’ performances on the post application of the achievement test showed a significant variance in favour of participants who received online teaching ( p < .05). Conclusion: The findings of the study suggest that, while both online and face-to-face mode can be effective in achieving learning goals of first aid content, online learning was found to be more effective in comparison to traditional classroom instruction.