bluuebunny/crossref_metadata_2025 huggingface dataset:
SNP Genotyping Defines Complex Gene-Flow Boundaries Among African Malaria Vector Mosquitoes
Authors: Neafsey D. E. , Lawniczak M. K. N. , Park D. J. , Redmond S. N. , Coulibaly M. B. , Traoré S. F. , Sagnon N. , Costantini C. , Johnson C. , Wiegand R. C. , Collins F. H. , Lander E. S. , Wirth D. F. , Kafatos F. C. , Besansky N. J. , Christophides G. K. , Muskavitch M. A. T.
Published: October 2010

Signals of Mosquito Speciation

Malaria in Africa is transmitted by the mosquito species complex
Anopheles gambiae
.

Neafsey
et al.

(p.
514
) made high-resolution single-nucleotide arrays to map genetic divergence among members of the species. Differentiation between populations was observed and evidence obtained for selective sweeps within populations. Most divergence occurred within inversion regions around the centrosome and in genes associated with development, pheromone signaling, and from the X chromosome. The analysis also revealed signals of sympatric speciation occurring within similar chromosomal regions in mosquitoes from different regions in Africa.

Lawniczak
et al.

(p.
512
) sequenced the genomes of two molecular forms (known as M and S) of
A. gambiae
, which have distinctive behavioral phenotypes and appear to be speciating. This effort resolves problems arising from the apparently chimeric nature of the reference genome and confirms the observed genome-wide divergences. This kind of analysis has the potential to contribute to control programs that can adapt to population shifts in mosquito behavior arising from the selective effects of the control measures themselves.
Politics and Parthenotes
Authors: Tingen C. , Rodriguez S. , Campo-Engelstein L. , Woodruff T. K.
Published: October 2010

Broad restrictions on funding for human embryo research can stymie work that was unnecessarily precluded by a flawed U.S. law.
Creating Ligands with Multiple Personalities
Authors: Crabtree Robert H.
Published: October 2010

Ligands are being designed to respond to the solution environment and tune the reactivity of the transition metals to which they bind.
Flipping the Light Switch
Authors: Vogt Nina , Desplan Claude
Published: October 2010

A specific neuronal circuit controls how flies respond to light during development.
The Flu's Proton Escort
Authors: Fiorin Giacomo , Carnevale Vincenzo , DeGrado William F.
Published: October 2010

A flurry of structural data provides sometimes conflicting insights into the M2 proton channel.
The Agora of Knowledge
WeltWissen [World Knowledge]
300 Jahre Wissenschaften in Berlin

Jochen Hennig, Curator

Martin-Gropius-Bau, Berlin. Through 9 January 2011. www.weltwissen-berlin.de
Authors: Laubichler Manfred D.
Published: October 2010

The exhibition's artifacts, installations, and media stations offer a variety of perspectives on the practice of science in Berlin over the past 300 years.
An Education That Inspires
Authors: Alberts Bruce
Published: October 2010

Why is it that children, who enter school at age 5 filled with excitement and wonder about the world, often become bored with education before their teenage years? How might the United States produce a more engaging education system, one that allows a child with a specific fascination to explore that interest in depth as an integral part of his or her early education? Here I sketch a possible plan based on science, technology, engineering, and math (STEM) awards that would be largely earned through student activities outside of school.
First Goldilocks Exoplanet May Not Exist
Authors: Kerr Richard A.
Published: October 2010

At a meeting last week in Turin, Italy, a group of exoplanet hunters announced that its observations show no sign of Gliese 581g, which only a couple of weeks earlier a rival group had announced was the long-sought Earth-like habitable planet.
How Wet the Moon? Just Damp Enough to Be Interesting
Authors: Kerr Richard A.
Published: October 2010

A series of reports of new water-related results from the moon beginning on page
463
of this week's issue of
Science
shows that the moon's recently discovered hydrosphere is nothing like Earth's watery regime, and many enigmas remain.
Rinderpest, Deadly for Cattle, Joins Smallpox as a Vanquished Disease
Authors: Normile Dennis
Published: October 2010

Rinderpest, an infectious disease that has decimated cattle and devastated their keepers for millennia, is gone. The United Nations Food and Agriculture Organization announced on 14 October in Rome that a 16-year eradication effort has succeeded and fieldwork has ended.
1.3 Billion Divided by 6.5 Million, and Watch That Floating Decimal
Authors: Hvistendahl Mara
Published: October 2010

China's unique solution to getting a handle on a swelling migrant, or "floating," population in this year's census—adding presumed residents to an actual head count—unsettles some demographers.
Immune Therapy Steps Up the Attack
Authors: Couzin-Frankel Jennifer
Published: October 2010

After years of trying, cancer researchers say they are finally having success enlisting the body's own defenses to destroy tumors.
Radio Astronomers Take Arms Against a Sea of Signals
Authors: Bhattacharjee Yudhijit
Published: October 2010

Beset by interference from telecommunications and radar, researchers are devising workarounds to preserve their view of the long-wavelength universe.
Influenza and Meningococcal Vaccinations Are Effective in Healthy Subjects Treated with the Interleukin-1β-Blocking Antibody Canakinumab: Results of an Open-Label, Parallel Group, Randomized, Single-Center Study
Authors: Chioato A. , Noseda E. , Felix S. D. , Stevens M. , Del Giudice G. , Fitoussi S. , Kleinschmidt A.
Published: October 2010

The objective of this study was to evaluate the efficacy of influenza and meningococcal vaccines in healthy subjects exposed to the anti-interleukin-1β (anti-IL-1β) monoclonal antibody canakinumab. This was an open-label, parallel group, randomized, single-center study of healthy subjects (aged 18 to 45 years). At baseline, antibody (Ab) titers were measured and subjects were randomized (1:1) to a single 300-mg canakinumab dose administered subcutaneously (s.c.) or received no treatment (control group). After 2 weeks, subjects were treated with inactivated, unadjuvanted influenza and conjugated group C meningococcal (MenC) vaccines, administered intramuscularly (i.m.). The primary efficacy variable was the response (≥2-fold increase in Ab titer in ≥2 of 3 influenza virus strains) after 4 weeks in subjects treated with canakinumab compared to the control group. Secondary efficacy variables were the antibody response to vaccines at different thresholds and time points. Fifty-one of 112 subjects screened were randomized to canakinumab (
n
= 25) or the control group (
n
= 26). Antibody responses to vaccinations measured against different influenza virus strains and one MenC strain at 4 weeks were comparable in the canakinumab and control groups. The primary efficacy variable, the response to influenza vaccination (≥2-fold increase in Ab titer in ≥2 of 3 serotypes) at 4 weeks, was shown in 24/25 subjects in the canakinumab group compared to 25/25 subjects in the control group. Antibody responses remained comparable in the two groups at the different time points assessed. Headache was the most frequently reported adverse event. No deaths or serious adverse events were reported during the study. We concluded that a single dose of 300 mg canakinumab s.c. does not affect the induction or persistence of antibody responses after vaccination with unadjuvanted influenza or alum-adjuvanted MenC vaccines in healthy subjects.
Contemporary Seasonal Influenza A (H1N1) Virus Infection Primes for a More Robust Response To Split Inactivated Pandemic Influenza A (H1N1) Virus Vaccination in Ferrets
Authors: Ellebedy Ali H. , Fabrizio Thomas P. , Kayali Ghazi , Oguin Thomas H. , Brown Scott A. , Rehg Jerold , Thomas Paul G. , Webby Richard J.
Published: October 2010

Human influenza pandemics occur when influenza viruses to which the population has little or no immunity emerge and acquire the ability to achieve human-to-human transmission. In April 2009, cases of a novel H1N1 influenza virus in children in the southwestern United States were reported. It was retrospectively shown that these cases represented the spread of this virus from an ongoing outbreak in Mexico. The emergence of the pandemic led to a number of national vaccination programs. Surprisingly, early human clinical trial data have shown that a single dose of nonadjuvanted pandemic influenza A (H1N1) 2009 monovalent inactivated vaccine (pMIV) has led to a seroprotective response in a majority of individuals, despite earlier studies showing a lack of cross-reactivity between seasonal and pandemic H1N1 viruses. Here we show that previous exposure to a contemporary seasonal H1N1 influenza virus and to a lesser degree a seasonal influenza virus trivalent inactivated vaccine is able to prime for a higher antibody response after a subsequent dose of pMIV in ferrets. The more protective response was partially dependent on the presence of CD8
+
cells. Two doses of pMIV were also able to induce a detectable antibody response that provided protection from subsequent challenge. These data show that previous infection with seasonal H1N1 influenza viruses likely explains the requirement for only a single dose of pMIV in adults and that vaccination campaigns with the current pandemic influenza vaccines should reduce viral burden and disease severity in humans.
Assessment of the IgA Immunoassay Diagnostic Potential of the
Mycobacterium tuberculosis
MT10.3-MPT64 Fusion Protein in Tuberculous Pleural Fluid
Authors: Araujo Leonardo Silva , Moraes Renata Maciel , Trajman Anete , Saad Maria Helena Féres
Published: October 2010

Pleural tuberculosis (PL-TB) remains difficult to diagnose. An enzyme-linked immunosorbent assay (ELISA) was developed based on a construction containing the fusion of the
Rv3019c
(MT10.3) and
Rv1980c
(MPT64) gene sequences, and its performance was evaluated in an area where TB is endemic. A total of 92 pleural fluid (PF) samples at serial dilutions of 1:50 to 1:800 were included in the ELISA IgA MT10.3-MPT64 evaluation: 70 from TB patients and 22 from patients with other pleurisies. Confirmation of the expression and subsequent purification of the protein was made by SDS-PAGE and Western blot assays, resulting in a 36-kDa protein. ELISA IgA MT10.3-MPT64 showed sensitivities of 61.4%, 58.6%, 62.9%, 67.1%, and 70% at each PF dilution, respectively. The cumulative results of all dilutions increased sensitivity to 81.4% without jeopardizing specificity. Similar results were also obtained at the combined dilutions of 1:50, 1:200, and 1:800 or 1:50 plus 1:800 dilutions (80%). The overall sensitivity of the reference test, i.e., histopathological examination, was 74%. But, via the ELISA IgA MT10.3-MPT64 test, sensitivity was high for specimens with a negative culture (23/27; 85.2%) or nonspecific histopathology (17/18; 94.4%). Our findings demonstrated the promising use of this test as an adjunct in PL-TB diagnoses, particularly in cases with lower bacterial loads and false-negative results in the reference tests, since the new test includes such important features as quick and easy application, high sensitivity and, perhaps most importantly, affordability, which is so crucial for its widespread use in developing countries.
Molecular Probes for Diagnosis of Clinically Relevant Bacterial Infections in Blood Cultures
Authors: Hansen Wendy L. J. , Beuving Judith , Bruggeman Cathrien A. , Wolffs Petra F. G.
Published: October 2010

Broad-range real-time PCR and sequencing of the 16S rRNA gene region is a widely known method for the detection and identification of bacteria in clinical samples. However, because of the need for sequencing, such identification of bacteria is time-consuming. The aim of our study was to develop a more rapid 16S real-time PCR-based identification assay using species- or genus-specific probes. The Gram-negative bacteria were divided into
Pseudomonas
species,
Pseudomonas aeruginosa
,
Escherichia coli
, and other Gram-negative species. Within the Gram-positive species, probes were designed for
Staphylococcus
species,
Staphylococcus aureus
,
Enterococcus
species,
Streptococcus
species, and
Streptococcus pneumoniae
. The assay also included a universal probe within the 16S rRNA gene region for the detection of all bacterial DNA. The assay was evaluated with a collection of 248 blood cultures. In this study, the universal probe and the probes targeting
Pseudomonas
spp.,
P. aeruginosa
,
E. coli
,
Streptococcus
spp.,
S. pneumoniae
,
Enterococcus
spp., and
Staphylococcus
spp. all had a sensitivity and specificity of 100%. The probe specific for
S. aureus
showed eight discrepancies, resulting in a sensitivity of 100% and a specificity of 93%. These data showed high agreement between conventional testing and our novel real-time PCR assay. Furthermore, this assay significantly reduced the time needed for identification. In conclusion, using pathogen-specific probes offers a faster alternative for pathogen detection and could improve the diagnosis of bloodstream infections.
Vancomycin Susceptibility Trends and Prevalence of Heterogeneous Vancomycin-Intermediate
Staphylococcus aureus
in Clinical Methicillin-Resistant
S. aureus
Isolates
Authors: Pitz Adam M. , Yu Fang , Hermsen Elizabeth D. , Rupp Mark E. , Fey Paul D. , Olsen Keith M.
Published: October 2010

Due to the rise in methicillin-resistant
Staphylococcus aureus
(MRSA) infections and widespread use of vancomycin, MRSA isolates with reduced susceptibility to vancomycin are emerging (i.e., MIC creep). However, the prevalence of heterogeneous vancomycin-intermediate
S. aureus
(hVISA) is unknown due to the difficulty in detecting this phenotype. Recently, Etest glycopeptide resistance detection (GRD) strips have been developed to detect hVISA. This study assessed vancomycin susceptibility in MRSA isolates and determined the prevalence of hVISA by Etest GRD and population analysis profile-area under the curve ratio (PAP-AUC). The genetic backgrounds of 167 MRSA isolates collected from 2000 to 2008 were identified by pulsed-field gel electrophoresis. Vancomycin MICs were determined using Etest and two broth microdilution assays, MicroScan and Sensititre. Etest GRD was performed on all isolates, and those exhibiting a hVISA phenotype were further tested by PAP-AUC. The vancomycin MIC modes remained consistent at 1 μg/ml, as assessed by Sensititre and MicroScan. Etest reported a significant increase (mode MIC = 1.5 μg/ml) in the MIC between 2000 and 2008 (
P
< 0.01); however, this increase did not reflect a ≥2-fold change. In addition, the slight MIC increase did not increase linearly from 2000 to 2008, suggesting biological fluctuation, and is inconsistent with the concept of MIC creep. Etest GRD identified six hVISA isolates, two of which were confirmed to be hVISA by PAP-AUC. In conclusion, reduced vancomycin susceptibility was not detected in our hospital over a 9-year period using three different MIC methodologies, and the hVISA incidence was 1.2%, as determined by Etest GRD and PAP-AUC.
Evaluation of a New Chromogenic Medium (StrepB
Select
) for Detection of Group B
Streptococcus
from Vaginal-Rectal Specimens
Authors: Louie L. , Kotowich L. , Meaney H. , Vearncombe M. , Simor A. E.
Published: October 2010

We compared StrepB
Select
medium (
Select
) after enrichment with conventional culture for the detection of Group B
Streptococcus
(GBS). Postenrichment sensitivities of
Select
and conventional culture were 98.8% and 92.2%, respectively (
P
< 0.05).
Select
was superior for detection of GBS from vaginal-rectal specimens. Growth of non-GBS colonies required additional work to exclude the presence of GBS, especially after 48 h of incubation. Incubation of
Select
beyond 24 h did not significantly increase the yield of GBS.
Evaluation of the BD GeneOhm Assay Using the Rotor-Gene 6000 Platform for Rapid Detection of Methicillin-Resistant
Staphylococcus aureus
from Pooled Screening Swabs
Authors: Smith Melvyn Howard , Hodgson Julian , Eltringham Ian Joseph
Published: October 2010

As health services move toward universal methicillin-resistant
Staphylococcus aureus
(MRSA) screening for hospital admissions, the most cost-effective approach is yet to be defined. In this study, one of the largest to date, we evaluated the performance of the BD GeneOhm MRSA assay on the Rotor-Gene 6000 thermal cycler, using samples taken directly from pooled MRSA screens. Results were compared with the same assay performed on the Smart-Cycler II platform and overnight broth culture. Samples yielding discrepant results were subjected to detailed analysis with an in-house PCR and patient note review. A total of 1,428 pooled MRSA screens were tested. Sensitivities and specificities of 85.3% and 95.8% for the Rotor-Gene and 81% and 95.7% for the Smart-Cycler were obtained, compared with broth enrichment. The sensitivity of the BD GeneOhm assay was increased to 100% when the results of in-house PCR and patient note review were taken into account. This study demonstrates that the Rotor-Gene 6000 thermal cycler is a reliable platform for use with the BD GeneOhm assay. It also proves that commercial PCR can be performed direct on pooled samples in selective broth, without the need for overnight incubation.
Molecular Detection of Mixed Infections of
Mycobacterium tuberculosis
Strains in Sputum Samples from Patients in Karonga District, Malawi
Authors: Mallard Kim , McNerney Ruth , Crampin Amelia C. , Houben Rein , Ndlovu Richard , Munthali Lumbani , Warren Robin M. , French Neil , Glynn Judith R.
Published: October 2010

The occurrence of mixed infections of
Mycobacterium tuberculosis
is no longer disputed. However, their frequency, and the impact they may have on our understanding of tuberculosis (TB) pathogenesis and epidemiology, remains undetermined. Most previous studies of frequency applied genotyping techniques to cultured
M. tuberculosis
isolates and found mixed infections to be rare. PCR-based techniques may be more sensitive for detecting multiple
M. tuberculosis
strains and can be applied to sputum. To date, one study in South Africa has used a PCR approach and suggested that mixed infection could be common. We investigated mixed infections in northern Malawi using two lineage-specific PCR assays targeting the Latin American-Mediterranean (LAM) and Beijing lineages. Compared with spoligotyping, the specificity and sensitivity of both assays was 100%. From 160 culture-positive sputa, mixed LAM and non-LAM strains were detected in 4 sputa belonging to 2 (2.8%) patients. Both patients were HIV positive, with no history of TB. Cultured isolates from both patients showed only LAM by PCR and spoligotyping. In a set of 377 cultured isolates, 4 were mixed LAM and non-LAM. Only one showed evidence of more than one
M. tuberculosis
strain using IS
6110
-based restriction fragment length polymorphism (IS
6110
-RFLP) and spoligotyping analyses. Corresponding sputa for the 4 isolates were unavailable. Mixed Beijing and non-Beijing strains were not detected in this study. Mixed infections appear to be rare in our setting and are unlikely to affect findings based on DNA fingerprinting data. Molecular methods, which avoid the selective nature of culture and target distinct strains, are well suited to detection of mixed infections.
False-Negative PCR Result Due to Gene Polymorphism: the Example of
Neisseria meningitidis
Authors: Jaton Katia , Ninet Béatrice , Bille Jacques , Greub Gilbert
Published: October 2010

Early treatment of meningococcal meningitis is mandatory but may negate the cerebrospinal fluid culture. Etiological diagnosis then mainly relies on PCR. Here, we report a case of false-negative results for real-time PCR for a
Neisseria meningitidis
serogroup B isolate with a polymorphism in the
ctrA
gene.
Blood Myeloid Dendritic Cells from HIV-1-Infected Individuals Display a Proapoptotic Profile Characterized by Decreased Bcl-2 Levels and by Caspase-3+Frequencies That Are Associated with Levels of Plasma Viremia and T Cell Activation in an Exploratory Study
Authors: Dillon Stephanie M. , Friedlander Laura J. , Rogers Lisa M. , Meditz Amie L. , Folkvord Joy M. , Connick Elizabeth , McCarter Martin D. , Wilson Cara C.
Published: October 2010

Reduced frequencies of myeloid and plasmacytoid dendritic cell (DC) subsets (mDCs and pDCs, respectively) have been observed in the peripheral blood of HIV-1-infected individuals throughout the course of disease. Accumulation of DCs in lymph nodes (LNs) may partly account for the decreased numbers observed in blood, but increased DC death may also be a contributing factor. We used multiparameter flow cytometry to evaluate pro- and antiapoptotic markers in blood mDCs and pDCs from untreated HIV-1-infected donors, from a subset of infected donors before and after receiving antiretroviral therapy (ART), and from uninfected control donors. Blood mDCs, but not pDCs, from untreated HIV-1-infected donors expressed lower levels of antiapoptotic Bcl-2 than DCs from uninfected donors. A subset of HIV-1-infected donors had elevated frequencies of proapoptotic caspase-3+blood mDCs, and positive correlations were observed between caspase-3+mDC frequencies and plasma viral load and CD8+T-cell activation levels. Caspase-3+mDC frequencies, but not mDC Bcl-2 expression, were reduced with viral suppression on ART. Apoptosis markers on DCs in blood and LN samples from a cohort of untreated, HIV-1-infected donors with chronic disease were also evaluated. LN mDCs displayed higher levels of Bcl-2 and lower caspase-3+frequencies than did matched blood mDCs. Conversely, LN pDCs expressed lower Bcl-2 levels than their blood counterparts. In summary, blood mDCs from untreated HIV-1-infected subjects displayed a proapoptotic profile that was partially reversed with viral suppression, suggesting that DC death may be a factor contributing to blood DC depletion in the setting of chronic, untreated HIV disease.
Yeast-Elicited Cross-Reactive Antibodies to HIV Env Glycans Efficiently Neutralize Virions Expressing Exclusively High-Mannose N-Linked Glycans
Authors: Agrawal-Gamse Caroline , Luallen Robert J. , Liu Bingfen , Fu Hu , Lee Fang-Hua , Geng Yu , Doms Robert W.
Published: October 2010

The HIV envelope (Env) protein uses a dense coat of glycans to mask conserved domains and evade host humoral immune responses. The broadly neutralizing antibody 2G12, which binds a specific cluster of high-mannose glycans on HIV Env, shows that the glycan shield can also serve as a target for neutralizing antibodies. We have described a triple mutant
Saccharomyces cerevisiae
strain that expresses high-mannose glycoproteins that bind to 2G12. When used to immunize rabbits, this yeast elicits antibodies that bind to gp120-associated glycans but fail to neutralize virus. Here we sought to determine the reason for these discordant results. Affinity purification of sera over columns conjugated with three 2G12-reactive yeast glycoproteins showed that these proteins could adsorb 80% of the antibodies that bind to gp120 glycans. Despite binding to monomeric gp120, these mannose-specific antibodies failed to bind cell surface-expressed trimeric Env. However, when Env was expressed in the presence of the mannosidase inhibitor kifunensine to force retention of high-mannose glycans at all sites, the purified antibodies gained the abilities to bind trimeric Env and to strongly and broadly neutralize viruses produced under these conditions. Combined, these data show that the triple mutant yeast strain elicits antibodies that bind to high-mannose glycans presented on the HIV envelope, but only when they are displayed in a manner not found on native Env trimers. This implies that the underlying structure of the protein scaffold used to present the high-mannose glycans may be critical to allow elicitation of antibodies that recognize trimeric Env and neutralize virus.
Small-Molecule Inhibition of Human Immunodeficiency Virus Type 1 Infection by Virus Capsid Destabilization
Authors: Shi Jiong , Zhou Jing , Shah Vaibhav B. , Aiken Christopher , Whitby Kevin
Published: October 2010

Human immunodeficiency virus type 1 (HIV-1) infection is dependent on the proper disassembly of the viral capsid, or “uncoating,” in target cells. The HIV-1 capsid consists of a conical multimeric complex of the viral capsid protein (CA) arranged in a hexagonal lattice. Mutations in CA that destabilize the viral capsid result in impaired infection owing to defects in reverse transcription in target cells. We describe here the mechanism of action of a small molecule HIV-1 inhibitor, PF-3450074 (PF74), which targets CA. PF74 acts at an early stage of HIV-1 infection and inhibits reverse transcription in target cells. We show that PF74 binds specifically to HIV-1 particles, and substitutions in CA that confer resistance to the compound prevent binding. A single point mutation in CA that stabilizes the HIV-1 core also conferred strong resistance to the virus without inhibiting compound binding. Treatment of HIV-1 particles or purified cores with PF74 destabilized the viral capsid
in vitro
. Furthermore, the compound induced the rapid dissolution of the HIV-1 capsid in target cells. PF74 antiviral activity was promoted by binding of the host protein cyclophilin A to the HIV-1 capsid, and PF74 and cyclosporine exhibited mutual antagonism. Our data suggest that PF74 triggers premature HIV-1 uncoating in target cells, thereby mimicking the activity of the retrovirus restriction factor TRIM5α. This study highlights uncoating as a step in the HIV-1 life cycle that is susceptible to small molecule intervention.
The Magnitude of Local Immunity in the Lungs of Mice Induced by Live Attenuated Influenza Vaccines Is Determined by Local Viral Replication and Induction of Cytokines
Authors: Lau Yuk-Fai , Santos Celia , Torres-Vélez Fernando J. , Subbarao Kanta
Published: October 2010

While live attenuated influenza vaccines (LAIVs) have been shown to be efficacious and have been licensed for human use, the surface glycoproteins hemagglutinin (HA) and neuraminidase (NA) have to be updated for optimal protective efficacy. Little is known about the effect of different HA and NA proteins on the immunogenicity of LAIVs developed using the same backbone. A panel of LAIVs that share the internal protein genes, with unique HA and NA gene segments from different influenza subtypes, was rescued by reverse genetics, and a comparative study of immune responses induced by these vaccines was conducted in mice. The results suggest that the magnitude of lung immunity, including pulmonary IgA antibody and memory CD8
+
T lymphocytes, induced by the vaccines depends on the replication efficiency of the LAIVs, as well as the induction of cytokines/chemokines in the lungs. However, these factors are not important in determining systemic immunity such as serum antibody titers and memory CD8
+
T cells in the spleen. A qualitative analysis of immune responses induced by a single dose of an H5N1 LAIV revealed that the vaccine induced robust systemic and mucosal immunity in mice. In addition, antibodies and memory lymphocytes established in the lungs following vaccination were required for protection against lethal challenge with homologous and heterologous H5N1 viruses. Our results highlight the different requirements for inducing systemic and lung immunity that can be explored for the development of pulmonary immunity for protection against respiratory pathogens.
Replicative and Transcriptional Activities of Hepatitis B Virus in Patients Coinfected with Hepatitis B and Hepatitis Delta Viruses
Authors: Pollicino Teresa , Raffa Giuseppina , Santantonio Teresa , Gaeta Giovanni Battista , Iannello Giuliano , Alibrandi Angela , Squadrito Giovanni , Cacciola Irene , Calvi Chiara , Colucci Giuseppe , Levrero Massimo , Raimondo Giovanni
Published: October 2010

Hepatitis B virus (HBV) and hepatitis delta virus (HDV) interplay was investigated by examining liver and serum samples from 21 coinfected and 22 HBV-monoinfected patients with chronic liver disease. Different real-time PCR assays were applied to evaluate intrahepatic amounts of HBV DNA, covalently closed circular DNA (cccDNA), pregenomic RNA (pgRNA), pre-S/S RNAs, and HDV RNA. Besides HBV DNA and HDV RNA levels, HBsAg concentrations in the sera were also determined. HDV-coinfected cases showed significantly lower median levels of serum HBV DNA (−5 log), intrahepatic relaxed-circular DNA (−2 log), and cccDNA (−2 log) than those of HBV-monoinfected cases. Interestingly, pgRNA and pre-S/S RNA amounts were significantly lower (both −1 log) in HDV-positive patients, whereas serum HBsAg concentrations were comparable between the two patient groups. Pre-S/S RNA and HBsAg amounts per cccDNA molecule were higher in HDV-positive patients (3-fold and 1 log, respectively), showing that HBV replication was reduced, whereas synthesis of envelope proteins was not specifically decreased. The ratios of cccDNA to intracellular total HBV DNA showed a larger proportion of cccDNA molecules in HDV-positive cases. For these patients, both intrahepatic and serum HDV RNA amounts were associated with cccDNA but not with HBsAg or HBV DNA levels. Finally, HBV genomes with large deletions in the basal core promoter/precore region were detected in 5/21 HDV-positive patients but in no HDV-negative patients and were associated with lower viremia levels. These findings provide significant information about the interference exerted by HDV on HBV replication and transcription activities in the human liver.
Induction of Type I Interferon Secretion through Recombinant Newcastle Disease Virus Expressing Measles Virus Hemagglutinin Stimulates Antibody Secretion in the Presence of Maternal Antibodies
Authors: Kim Dhohyung , Martinez-Sobrido Luis , Choi Changsun , Petroff Natasha , García-Sastre Adolfo , Niewiesk Stefan , Carsillo Thomas
Published: October 2010

Measles virus (MV) vaccine effectively protects seronegative individuals against infection. However, inhibition of vaccine-induced seroconversion by maternal antibodies after vaccination remains a problem, as it leaves infants susceptible to MV infection. In cotton rats, passive transfer of MV-specific IgG mimics maternal antibodies and inhibits vaccine-induced seroconversion. Here, we report that immunization in the presence of passively transferred IgG inhibits the secretion of neutralizing antibodies but not the generation of MV-specific B cells. This finding suggested that MV-specific B cells require an additional stimulus to mature into antibody-secreting plasma cells. In order to provide such a stimulus, we generated a recombinant Newcastle disease virus (NDV) expressing the MV hemagglutinin (NDV-H). In contrast to MV, NDV-H induced high levels of type I interferon in plasmacytoid dendritic cells and in lung tissue. In cotton rats immunized with NDV-H, neutralizing antibodies were also generated in the presence of passively transferred antibodies. In the latter case, however, the level and kinetics of antibody generation were reduced.In vitro, alpha interferon stimulated the activation of MV-specific B cells from MV-immune spleen cells. NDV infection (which induces alpha interferon) had the same effect, and stimulation could be abrogated by antibodies neutralizing alpha interferon, but not interleukin 6 (IL-6).In vivo, coapplication of UV-inactivated MV with NDV led to increased MV-specific antibody production in the presence and absence of passively transferred antibodies. These data indicate that MV-specific B cells are being generated after immunization in the presence of maternal antibodies and that the provision of alpha interferon as an additional signal leads to antibody secretion.
Poliovirus-Mediated Disruption of Cytoplasmic Processing Bodies
Authors: Dougherty Jonathan D. , White James P. , Lloyd Richard E.
Published: October 2010

Metazoan cells form cytoplasmic mRNA granules such as stress granules (SG) and processing bodies (P bodies) that are proposed to be sites of aggregated, translationally silenced mRNAs and mRNA degradation. Poliovirus (PV) is a plus-strand RNA virus containing a genome that is a functional mRNA; thus, we investigated if PV antagonizes the processes that lead to formation of these structures. We have previously shown that PV infection inhibits the ability of cells to form stress granules by cleaving RasGAP-SH3-binding protein (G3BP). Here, we show that P bodies are also disrupted during PV infection in cells by 4 h postinfection. The disruption of P bodies is more rapid and more complete than disruption of stress granules. The kinetics of P body disruption correlated with production of viral proteinases and required substantial viral gene product expression. The organizing mechanism that forms P body foci in cells is unknown; however, potential scaffolding, aggregating, or other regulatory proteins found in P bodies were investigated for degradation. Two factors involved in 5′-end mRNA decapping and degradation, Xrn1 and Dcp1a, and the 3′ deadenylase complex component Pan3 underwent accelerated degradation during infection, and Dcp1a may be a direct substrate of PV 3C proteinase. Several other key factors proposed to be essential for P body formation, GW182, Edc3, and Edc4, were unaffected by poliovirus infection. Since deadenylation has been reported to be required for P body formation, viral inhibition of deadenylation, through Pan3 degradation, is a potential mechanism of P body disruption.
Identification and Structural Characterization of the ALIX-Binding Late Domains of Simian Immunodeficiency Virus SIV
mac239
and SIV
agmTan-1
Authors: Zhai Qianting , Landesman Michael B. , Robinson Howard , Sundquist Wesley I. , Hill Christopher P.
Published: October 2010

Retroviral Gag proteins contain short late-domain motifs that recruit cellular ESCRT pathway proteins to facilitate virus budding. ALIX-binding late domains often contain the core consensus sequence YPX
n
L (where X
n
can vary in sequence and length). However, some simian immunodeficiency virus (SIV) Gag proteins lack this consensus sequence, yet still bind ALIX. We mapped divergent, ALIX-binding late domains within the p6
Gag
proteins of SIV
mac239
(
40
SREK
P
YKE
VT
ED
L
LHLNSLF
59
) and SIV
agmTan-1
(
24
AAG
A
YDP
AR
KL
L
EQYAKK
41
). Crystal structures revealed that anchoring tyrosines (in lightface) and nearby hydrophobic residues (underlined) contact the ALIX V domain, revealing how lentiviruses employ a diverse family of late-domain sequences to bind ALIX and promote virus budding.
Variola and Monkeypox Viruses Utilize Conserved Mechanisms of Virion Motility and Release That Depend on Abl and Src Family Tyrosine Kinases
Authors: Reeves Patrick M. , Smith Scott K. , Olson Victoria A. , Thorne Steve H. , Bornmann William , Damon Inger K. , Kalman Daniel
Published: October 2010

Vaccinia virus (VacV) enters mammalian cells, replicates extranuclearly, and produces virions that move to the cell surface along microtubules, fuse with the plasma membrane, and move from infected cells toward apposing cells on actin-filled membranous protrusions or actin tails. To form actin tails, cell-associated enveloped virions (CEV) require Abl and Src family tyrosine kinases. Furthermore, release of CEV from the cell requires Abl but not Src family tyrosine kinases and is blocked by imatinib mesylate (STI-571; Gleevec), an Abl family kinase inhibitor used to treat chronic myelogenous leukemia in humans. Here we demonstrate that the
Poxviridae
family members monkeypox virus (MPX) and variola virus (VarV) use conserved mechanisms for actin motility and extracellular enveloped virion (EEV) release. Furthermore, we show that imatinib mesylate is effective in a mouse model of infection with VacV, whether delivered prophylactically or postinfection, and restricts spread of virions from the site of inoculation. While inhibitors of both Src and Abl family kinases, such as dasatinib (BMS-354825; Sprycel), are effective in limiting dissemination of VacV, VarV, and MPX
in vitro
, members of this class of drugs appear to have immunosuppressive effects
in vivo
that preclude their use as anti-infectives. Together, these data suggest a possible utility for imatinib mesylate in treating smallpox or MPX infections or complications associated with vaccination.
Passive flooding of paranasal sinuses and middle ears as a method of equalisation in extreme breath-hold diving
Authors: Germonpré Peter , Balestra Costantino , Musimu Patrick
Published: October 2010

Breath-hold diving is both a recreational activity, performed by thousands of enthusiasts in Europe, and a high-performance competitive sport. Several ‘disciplines’ exist, of which the ‘no-limits’ category is the most spectacular: using a specially designed heavy ‘sled,’ divers descend to extreme depths on a cable, and then reascend using an inflatable balloon, on a single breath. The current world record for un-assisted descent stands at more than 200 m of depth. Equalising air pressure in the paranasal sinuses and middle-ear cavities is a necessity during descent to avoid barotraumas. However, this requires active insufflations of precious air, which is thus unavailable in the pulmonary system. The authors describe a diver who, by training, is capable of allowing passive flooding of the sinuses and middle ear with (sea) water during descent, by suppressing protective (parasympathetic) reflexes during this process. Using this technique, he performed a series of extreme-depth breath-hold dives in June 2005, descending to 209 m of sea water on one breath of air.
Referrals from a primary care-based sports medicine department to an orthopaedic department: a retrospective cohort study
Authors: Menzies Robert D , Young Richard A
Published: October 2010

Objective

To describe the impact of an expanded primary care-based sports medicine clinic on referrals to an orthopaedics clinic and to describe the patients seen and procedures performed.

Design

Retrospective cohort study.

Setting

Primary care-based sports medicine clinic and orthopaedics clinic at a tax-supported American safety net healthcare system.

Participants

All patients referred to the sports medicine clinic by other primary care physicians over a 1-year time period of July 2006–June 2007.

Main outcome measures

The referral rate from sports medicine clinic to orthopaedics clinic, the percentage of referred patients who were recommended surgery by the orthopaedists, the change in average waiting time to be seen in orthopaedics clinic and the most common conditions and procedures.

Results

4925 patients were seen by the sports medicine department; 118 (2.4%) of those patients were referred to the orthopaedic department. Of the referred patients, surgery was offered by orthopaedists to 80 (68%) patients. The average wait for initial consultation by the orthopaedic spine clinic decreased from 199 to 70 days; the wait for general orthopaedic clinic decreased from 97 to 19 days. No single patient complaint or musculoskeletal pathology predominated: knee degenerative joint disease (25.3%), mechanical low back pain (21.6%) and lumbar disc disease (19.9%). Knee injections and epidural steroid injections were the most common procedures performed.

Conclusions

Very few patients with musculoskeletal pathology were referred by a primary care-based sports medicine clinic to an orthopaedics clinic. Of the referred patients, sports medicine physicians and orthopaedists frequently agreed on the need for surgery. Expansion of a primary care-based sports medicine service could help relieve overburdened orthopaedics departments of patients with conditions not requiring surgery.
Clustered metabolic risk and leisure-time physical activity in adolescents: effect of dose?
Authors: Pahkala Katja , Heinonen Olli J , Lagström Hanna , Hakala Paula , Hakanen Maarit , Hernelahti Miika , Ruottinen Soile , Sillanmäki Lauri , Rönnemaa Tapani , Viikari Jorma , Raitakari Olli T , Simell Olli
Published: October 2010

Objectives

The authors studied the association of leisure-time physical activity (LTPA) with clustered and individual metabolic risk factors in adolescents taking into account diet and pubertal status. The authors also studied whether screen time was associated with clustered risk.

Methods

Self-reported LTPA and screen time, lipids, lipoproteins, apolipoproteins, high-sensitivity C reactive protein, blood pressure, body mass index (BMI), pubertal status and diet were assessed in 13-year-old adolescents (n=542) participating in an atherosclerosis prevention study (Special Turku Coronary Risk Factor Intervention Project for Children). Activity groups were formed according to sex-specific LTPA index tertile cut-off points. BMI, high-density lipoprotein cholesterol (HDL-C), triglycerides and blood pressure comprised the cluster.

Results

An increase in LTPA was associated with a decreased risk for clustered metabolic risk in girls. When sedentary and highly active adolescents were compared, an increase in LTPA decreased clustering of risk factors in boys as well. Little extra benefit on clustered risk was obtained by increasing LTPA from 30 MET h/week (eg, 4–5 h/week bicycling or playing soccer) to 50 MET h/week (eg, 7–8 h/week bicycling or playing soccer). LTPA was beneficially associated with BMI, HDL-C, systolic blood pressure and HDL-C/total cholesterol in girls and HDL-C in boys. Diet and pubertal status were similar in all activity groups. In girls, screen time >2 h/day was associated with an increased risk for clustered risk, independent of LTPA.

Conclusion

Sedentary adolescents had an increased risk for clustered metabolic risk compared with physically more active peers. Only minor extra benefit was obtained when LTPA increased over 30 MET h/week. Focus in the prevention of clustered risk should especially be on avoiding sedentary lifestyle.
The epidemiology of injuries in English youth community rugby union
Authors: Haseler Christine M , Carmont Michael R , England Michael
Published: October 2010

Introduction

Recent studies report the incidence and epidemiology of injury in professional rugby union; however, there is limited research in amateur and youth rugby. Injuries in youth rugby may have consequences for sports participation and physical development. The authors performed a prospective cohort study of injuries during youth community rugby.

Methods

An injury surveillance programme was established for the 2008–2009 season (9 months, 1636 player-hours) of an English community rugby club. The study included 210 players, all males, in Under 9 to Under 17 (U9–U17) age groups. These were categorised into mini, junior, pubertal and school participation age groupings. Injuries were defined according to the International Rugby Board consensus statements.

Results

There were 39 injuries reported (overall injury rate 24/1000 player-hours). Injury rates ranged from 0 to 49.3/1000 player-hours. More injuries occurred in junior (34.2/1000 player-hours) than in minis (11.9/1000 player-hours) (p<0.025). Higher numbers of moderate (20.6/1000 player-hours, p<0.005) and severe (9.5/1000 player-hours, p<0.05) injuries occurred in the U16–U17 age groups compared with younger age groups (U9–U10) where only minor injuries were reported. Most injuries occurred in the tackle (59%). The knee (4.9/1000 player-hours), shoulder (4.9/1000 player-hours) and head (4.3/1000 player-hours) were the most commonly affected areas. Concussion (1.8/1000 player-hours) affected half of the head injuries.

Conclusions

Injuries in youth rugby occur infrequently and are lower than in adult series. The risk of injury and severity of injury increases with age. This study highlights the need for further research into injury risk factors around puberty and the need for first aid provision.
Injury risk of playing football in Futsal World Cups
Authors: Junge Astrid , Dvorak Jiri
Published: October 2010

Background

While injuries in football (soccer) have been the subject of numerous publications, little information is available about the injury risk in futsal.

Objective

Analysis of the incidence and characteristics of injury in futsal players during top-level international tournaments.

Design

Prospective survey.

Methods

Player injuries during three consecutive Futsal World Cups were analysed using an established injury report system. The physicians of all participating teams reported all injuries after each match on a standardised injury report form. The average response rate was 93%.

Results

A total of 165 injuries were reported from 127 matches, which is equivalent to an incidence of 195.6 injuries per 1000 player hours (95% CI 165.8 to 225.6) or 130.4 injuries per 1000 player matches (95% CI 110.5 to 150.3). The majority of injuries were caused by contact with another player, and 36% of the injuries occurred during non-contact activities. Most injuries affected the lower extremity (70%), followed by head and neck (13%), upper extremity (10%) and trunk (7%). The most frequent diagnoses were contusion of the lower leg (11%), ankle sprain (10%) and groin strain (8%). On average, one time-loss injury in every two matches was reported.

Conclusion

While the location and diagnosis of injuries were similar in the two types of football, fewer injuries in futsal than in football were caused by contact with another player and by foul play. More information on injury mechanisms in futsal is needed to develop specific injury preventive interventions.
Clinicopathological evaluation of 100 cases of mucinous carcinoma of breast with emphasis on axillary staging and special reference to a micropapillary pattern
Authors: Ranade Aditi , Batra Rashmi , Sandhu Gagangeet , Chitale Rohit A , Balderacchi Jasminka
Published: October 2010

Background

Pure mucinous breast carcinoma (PMBC) is uncommon and associated with better prognosis than mixed mucinous breast carcinoma (MMBC). A micropapillary pattern in PMBC has been identified although its prognostic significance is questionable.

Methods

A retrospective review of 100 cases of mucinous carcinoma diagnosed between 2000 and 2009 was conducted. Two broad categories were studied: PMBC (more than 90% mucinous component; n=45) and MMBC (less than 90% mucinous component; n=55). PMBC was further subclassified as hypocellular/type A (n=37) and cellular/type B (n=8). Receptor status, clinicomorphological and prognostic features were compared without patient follow-up.

Results

Mean age at diagnosis in PMBC and MMBC was 60 and 63 years, while mean tumour size was 1.65 and 2.5 cm, respectively. Mean age in type A and type B PMBC patients was 75 and 55 years, respectively. The majority of PMBCs were well differentiated, with two poorly differentiated cases as well. The majority of MMBCs were moderately differentiated. A micropapillary pattern was seen in 20% of PMBCs. Sentinel lymph nodes were positive in 18.5% of PMBCs and 16% of MMBCs. Non-sentinel lymph nodes were positive in 14% of PMBCs and 39% of MMBCs. A micropapillary pattern was seen in 60% of LN positive PMBCs and 14% of LN negative PMBCs. Furthermore, 95% of PMBCs were ER(+), 84% were PR(+) and 9% were Her-2(+); 91% of MMBCs were ER(+), 87% were PR(+) and 33% were Her-2(+).

Conclusions

PMBCs with a micropapillary pattern were more frequently associated with nodal disease. PMBCs with axillary disease had one or more of the following: micropapillary pattern, high nuclear grade, Her-2 positivity, smaller tumour size or younger age. Hence, axillary staging by sentinel lymph node biopsy is recommended in PMBCs.