bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of Reeling on Fatigue Crack Growth Behavior of Welded API5LX65 Line Pipe
Authors: Thodla Ramgopal , Gordon Robin , Gui Feng
Published: October 2015

The effect of reeling on the fatigue crack growth rate (FCGR) behavior of welded pipe was investigated both in-air as well as in sour environment. The FCGR behavior of the reeled pipe in various notch locations parent pipe (PP), weld center line (WCL), heat affected zone (HAZ) did not exhibit any effect of reeling (i.e. the properties in the strained and aged condition was similar to the as-fabricated welds). Frequency scan FCGR tests in sour environment (pH = 5/0.46psia H2S) exhibited maximum KDF’s in the range of 10× to 35× at frequencies in the range of 3 mHz to 1 mHz and 3× to 5× at frequencies in the range of 0.3Hz (Risers). In sour service WCL exhibited better fatigue performance than the parent pipe and HAZ in all conditions. Fatigue performance of parent pipe and WCL was independent of reeling. The poorest fatigue performance was observed in unstrained HAZ. Fatigue performance of HAZ extrados and intrados were similar and better than unstrained HAZ. It was also found that the FCGR in sour environments was controlled by the internal hydrogen due to bulk charging from the sour environment. The overall conclusion is that, reeling has no detrimental effect on sour service fatigue crack growth behavior i.e. sour service fatigue performance of reeled pipe is the same as unreeled pipe.
Effect of Reeling on Fracture Toughness Behavior of Welded API5LX65 Line Pipe
Authors: Thodla Ramgopal , Gordon Robin , Gui Feng
Published: October 2015

The effect of reeling on the fracture toughness (FT) behavior of welded pipe was investigated both in-air as well as in sour environment. The FT behavior of the reeled pipe in various notch locations parent pipe (PP), heat affected zone (HAZ), weld center line (WCL) did not exhibit any effect of reeling (i.e. the properties in the strained and aged condition was similar to the as-fabricated welds). Fracture toughness in sour environments is 4× to 6× lower than the In-Air values. The fracture toughness behavior of parent pipe in sour environments was affected by reeling at strains associated with 2.12%/cycle and 2.72%/cycle. WCL fracture toughness was not affected by reeling in sour environments. The intrados HAZ exhibited the lowest fracture toughness properties. Some of the sour service fracture toughness tests on parent pipe and HAZ strained to 2.72%/cycle on the intrados samples exhibited pop-in behavior possibly due to higher YS. Parent pipe strained to a lower level of 2.12%/cycle did not exhibit any evidence of pop-in’s. Reeling appears to decrease the parent pipe toughness in the extrados and intrados in sour service both at 2.12%/cycle and 2.72%/cycle. The extrados parent pipe and intrados HAZ showed the lowest fracture toughness values in sour environment.
A Real-Time System for Forecasting Extreme Waves and Vessel Motions
Authors: Alford Laura K. , Beck Robert F. , Johnson Joel T. , Lyzenga David , Nwogu Okey , Zundel Alan
Published: October 2015

The University of Michigan is leading a team that includes subcontractors Ohio State University, Aquaveo, LLC, and Woods Hole Oceanographic Institute to design, implement, and test an Environmental and Ship Motion Forecasting (ESMF) system. The system has application to many challenges associated with offshore operations, including skin-to-skin transfer of cargo/personnel and extreme wave/response prediction. Briefly, the system uses a modified commercial-off-the-shelf (COTS) Doppler marine radar to determine the wave field surrounding the vessel; nonlinear wave theory to propagate the wave surface forward in time; and seakeeping theory to predict future vessel motions. A major challenge is that all computations must be done in real time. This paper will briefly describe the system and show an example application of predicting extreme waves and motions for a floating offshore type platform.
Fabrication of nanoporous membranes for tuning microbial interactions and biochemical reactions
Authors: Shankles Peter G. , Timm Andrea C. , Doktycz Mitchel J. , Retterer Scott T.
Published: October 2015

New strategies for combining conventional photo- and soft-lithographic techniques with high-resolution patterning and etching strategies are needed in order to produce multiscale fluidic platforms that address the full range of functional scales seen in complex biological and chemical systems. The smallest resolution required for an application often dictates the fabrication method used. Micromachining and micropowder blasting yield higher throughput, but lack the resolution needed to fully address biological and chemical systems at the cellular and molecular scales. In contrast, techniques such as electron beam lithography or nanoimprinting allow nanoscale resolution, but are traditionally considered costly and slow. Other techniques such as photolithography or soft lithography have characteristics between these extremes. Combining these techniques to fabricate multiscale or hybrid fluidics allows fundamental biological and chemical questions to be answered. In this study, a combination of photolithography and electron beam lithography are used to produce two multiscale fluidic devices that incorporate porous membranes into complex fluidic networks in order to control the flow of energy, information, and materials in chemical form. In the first device, materials and energy were used to support chemical reactions. A nanoporous membrane fabricated with e-beam lithography separates two parallel, serpentine channels. Photolithography was used to pattern microfluidic channels around the membrane. The pores were written at 150 nm and reduced in size with silicon dioxide deposition from plasma enhanced chemical vapor deposition and atomic layer deposition. Using this method, the molecular weight cutoff of the membrane can be adapted to the system of interest. In the second approach, photolithography was used to fabricate 200 nm thin pores. The pores confined microbes and allowed energy replenishment from a media perfusion channel. The same device can be used for study of intercellular communication via the secretion and uptake of signal molecules. Pore size was tested with 750 nm fluorescent polystyrene beads and fluorescein dye. The 200 nm polydimethylsiloxane pores were shown to be robust enough to hold 750 nm beads while under pressure, but allow fluorescein to diffuse across the barrier. Further testing showed that extended culture of bacteria within the chambers was possible. These two examples show how lithographically defined porous membranes can be adapted to two unique situations and used to tune the flow of chemical energy, materials, and information within a microfluidic network.
Immobilization of inorganic pyrophosphatase on nanodiamond particles retaining its high enzymatic activity
Authors: Rodina Elena V. , Valueva Anastasiya V. , Yakovlev Ruslan Yu. , Vorobyeva Nataliya N. , Kulakova Inna I. , Lisichkin Georgy V. , Leonidov Nikolay B.
Published: October 2015

Nanodiamond (ND) particles are popular platforms for the immobilization of molecular species. In the present research, enzyme Escherichia coli inorganic pyrophosphatase (PPase) was immobilized on detonation ND through covalent or noncovalent bonding and its enzymatic activity was characterized. Factors affecting adsorption of PPase such as ND size and surface chemistry were studied. The obtained material is a submicron size association of ND particles and protein molecules in approximately equal amounts. Both covalently and noncovalently immobilized PPase retains a significant enzymatic activity (up to 95% of its soluble form) as well as thermostability. The obtained hybrid material has a very high enzyme loading capacity (∼1 mg mg−1) and may be considered as a promising delivery system of biologically active proteinaceous substances, particularly in the treatment of diseases such as calcium pyrophosphate crystal deposition disease and related pathologies. They can also be used as recoverable heterogeneous catalysts in the traditional uses of PPase.
Effects of instructed timing and tempo on snare drum sound in drum kit performance
Authors: Danielsen Anne , Waadeland Carl Haakon , Sundt Henrik G. , Witek Maria A. G.
Published: October 2015

This paper reports on an experiment investigating the expressive means with which performers of groove-based musics signal the intended timing of a rhythmic event. Ten expert drummers were instructed to perform a rock pattern in three different tempi and three different timing styles: “laid-back,” “on-the-beat,” and “pushed.” The results show that there were systematic differences in the intensity and timbre (i.e., sound-pressure level, temporal centroid, and spectral centroid) of series of snare strokes played with these different timing styles at the individual level. A common pattern was found across subjects concerning the effect of instructed timing on sound-pressure level: a majority of the drummers played laid-back strokes louder than on-the-beat strokes. Furthermore, when the tempo increased, there was a general increase in sound-pressure level and a decrease in spectral centroid across subjects. The results show that both temporal and sound-related features are important in order to indicate that a rhythmic event has been played intentionally early, late, or on-the-beat, and provide insight into the ways in which musicians communicate at the microrhythmic level in groove-based musics.
Cardiovascular and Metabolic Responses to On-Water Upwind Sailing in Optimist Sailors
Authors: Lopez Santiago , Bourgois Jan G. , Tam Enrico , Bruseghini Paolo , Capelli Carlo
Published: October 2015

Purpose:

To explore the cardiovascular and metabolic responses of 9 Optimist sailors (12.7 ± 0.8 y, 153 ± 9 cm, 41 ± 6 kg, sailing career 6.2 ± 1 y, peak oxygen uptake [V̇O2peak] 50.5 ± 4.5 mL · min−1 · kg−1) during on-water upwind sailing with various wind intensities (W).

Methods:

In a laboratory session, peak V̇O2, beat-by-beat cardiac output (Q̇), mean arterial blood pressure (MAP), and heart rate (fH) were measured using a progressive cycle ramp protocol. Steady-state V̇O2, Q̇, MAP, and fH at 4 submaximal workloads were also determined. During 2 on-water upwind sailing tests (constant course and with tacks), W, Q̇, MAP, and fH were measured for 15 min. On-water V̇O2 was estimated on the basis of steady-state fH measured on water and of the individual ΔV̇O2/ΔfH relationship obtained in the laboratory.

Results:

V̇O2, fH, and Q̇ expressed as percentage of the corresponding peak values were linearly related with W; exercise intensity during on-water sailing corresponded to 46–48% of V̇O2peak. MAP and total vascular peripheral resistance (TPR = MAP/Q̇) were larger (P < .005) during on-water tests (+39% and +50%, respectively) than during cycling, and they were correlated with W. These responses were responsible for larger values of the double (DP) and triple (TP) products of the heart during sailing than during cycling (P < .005) (+37% and +32%, respectively).

Conclusions:

These data indicate that the cardiovascular system was particularly stressed during upwind sailing even though the exercise intensity of this activity was not particularly high.
Effect of Performance Level on the Prediction of Middle-Distance-Running Performances Using a Nomogram
Authors: Tabben Montassar , Bosquet Laurent , Coquart Jeremy B.
Published: October 2015

Purpose:

This study examined the effect of performance level on the validity and accuracy of middle-distance running-performance predictions obtained from the nomogram of Mercier et al in male runners.

Methods:

Official French track-running rankings for the 3000-, 5000-, and 10,000-m events from 2006 to 2014 were examined. The performance level was determined from the official reference table of the Fédération Française d’Athlétisme, and the runners were divided in 3 groups (ie, low, moderate, and high levels). Only male runners who performed in the 3 distance events within the same year were included (N = 443). Each performance over any distance was predicted using the nomogram from the 2 other performances.

Results:

No difference was found in low- and moderate-performance-level athletes (0.02 ≤ effect size [ES] ≤ 0.06, 95% limits of agreement [LoA] ≤ 6%). By contrast, a small difference in high-performance-level athletes (P < .01, 0.23 ≤ ES ≤ 0.45, 95% LoA ≤ 11.6%) was found.

Conclusion:

The study confirms the validity of the nomogram to predict track-running performance with a high level of accuracy, except for male runners with high performance level (ie, national or international). Consequently, the predictions from the nomogram may be used in training programs (eg, to prescribe tempo runs with realistic training velocities) and competitions (eg, to plan realistic split times to reach the best performance).
The Effect of Mindfulness Training on Attention and Performance in National-Level Swimmers: An Exploratory Investigation
Authors: Mardon Nick , Richards Hugh , Martindale Amanda
Published: October 2015

This quasi-experimental intervention study investigated the impact of mindfulness training on attention and performance in swimmers. Following an 8-week intervention with six national-level university swimmers (M = 20 years), single case analysis of pre- and post- measurements for three of six participants showed large improvements in mindfulness and attention efficiency. Two participants showed a small increase in one of mindfulness or attention efficiency, and one showed no changes. Four participants improved performance times compared with season-best, and five participants improved self-rated performance. Athletes and coach positively evaluated mindfulness training. This study, with strong ecological validity, shows improvements in mindfulness, attention, and performance, consistent with theory that proposes attention as a mechanism for mindfulness based performance changes. Mindfulness training can be an effective and practical intervention. Further applied research is required utilizing designs to determine causality and further test the proposed mechanisms through which mindfulness may influence performance.
Immune correlates of vaccine protection against HIV-1 acquisition
Authors: Corey Lawrence , Gilbert Peter B. , Tomaras Georgia D. , Haynes Barton F. , Pantaleo Giuseppe , Fauci Anthony S.
Published: October 2015

Vaccine-induced non–broadly neutralizing antibodies are associated with protection against HIV-1 acquisition.
Neonatal Pasteurella multocida subsp. septica Meningitis Traced to Household Cats: Molecular Linkage Analysis Using Repetitive-Sequence-Based PCR
Authors: Boyanton Bobby L. , Freij Bishara J. , Robinson-Dunn Barbara , Makin Jacob , Runge Jessica K. , Luna Ruth Ann
Published: October 2015

Pasteurella multocida
is a rare cause of neonatal bacterial meningitis. We describe such a case and verify two household cats as the source of infection using repetitive-element PCR (rep-PCR) molecular fingering.
Multicenter Evaluation of Anyplex Plus MTB/NTM MDR-TB Assay for Rapid Detection of Mycobacterium tuberculosis Complex and Multidrug-Resistant Isolates in Pulmonary and Extrapulmonary Specimens
Authors: Sali Michela , De Maio Flavio , Caccuri Francesca , Campilongo Federica , Sanguinetti Maurizio , Fiorentini Simona , Delogu Giovanni , Giagulli Cinzia
Published: October 2015

The rapid diagnosis of tuberculosis (TB) and the detection of drug-resistant
Mycobacterium tuberculosis
strains are critical for successful public health interventions. Therefore, TB diagnosis requires the availability of diagnostic tools that allow the rapid detection of
M. tuberculosis
and drug resistance in clinical samples. Here, we performed a multicenter study to evaluate the performance of the Seegene Anyplex MTB/NTM MDR-TB assay, a new molecular method based on a multiplex real-time PCR system, for detection of
Mycobacterium tuberculosis
complex (MTBC), nontuberculous mycobacteria (NTM), and genetic determinants of drug resistance. In total, the results for 755 samples (534 pulmonary and 221 extrapulmonary samples) were compared with the results of smears and cultures. For pulmonary specimens, the sensitivities of the Anyplex assay and acid-fast bacillus smear testing were 86.4% and 75.0%, respectively, and the specificities were 99% and 99.4%. For extrapulmonary specimens, the sensitivities of the Anyplex assay and acid-fast bacillus smear testing were 83.3% and 50.0%, respectively, and the specificities of both were 100%. The negative and positive predictive values of the Anyplex assay for pulmonary specimens were 97% and 100%, respectively, and those for extrapulmonary specimens were 84.6% and 100%. The sensitivities of the Anyplex assay for detecting isoniazid resistance in MTBC strains from pulmonary and extrapulmonary specimens were 83.3% and 50%, respectively, while the specificities were 100% for both specimen types. These results demonstrate that the Anyplex MTB/NTM MDR-TB assay is an efficient and rapid method for the diagnosis of pulmonary and extrapulmonary TB and the detection of isoniazid resistance.
Clinical and Laboratory Testing for Trichomonas vaginalis Infection
Authors: Van Der Pol Barbara
Published: October 2015

Trichomonas vaginalis
infection is highly prevalent in the United States and worldwide. Traditional clinical diagnostic methods fail to identify more than half of these infections that, if left untreated, can result in adverse pregnancy outcomes and an exacerbated risk of both acquisition and transmission of HIV. Women bear a disproportionate amount of the burden of these infections, and testing among populations at risk for this disease should be provided. Molecular technologies have expanded our capacity for laboratory-based detection of infection and can be used on samples already being collected for chlamydia/gonorrhea screening.
Development of a TaqMan Array Card for Acute-Febrile-Illness Outbreak Investigation and Surveillance of Emerging Pathogens, Including Ebola Virus
Authors: Liu Jie , Ochieng Caroline , Wiersma Steve , Ströher Ute , Towner Jonathan S. , Whitmer Shannon , Nichol Stuart T. , Moore Christopher C. , Kersh Gilbert J. , Kato Cecilia , Sexton Christopher , Petersen Jeannine , Massung Robert , Hercik Christine , Crump John A. , Kibiki Gibson , Maro Athanasia , Mujaga Buliga , Gratz Jean , Jacob Shevin T. , Banura Patrick , Scheld W. Michael , Juma Bonventure , Onyango Clayton O. , Montgomery Joel M. , Houpt Eric , Fields Barry
Published: October 2015

Acute febrile illness (AFI) is associated with substantial morbidity and mortality worldwide, yet an etiologic agent is often not identified. Convalescent-phase serology is impractical, blood culture is slow, and many pathogens are fastidious or impossible to cultivate. We developed a real-time PCR-based TaqMan array card (TAC) that can test six to eight samples within 2.5 h from sample to results and can simultaneously detect 26 AFI-associated organisms, including 15 viruses (chikungunya, Crimean-Congo hemorrhagic fever [CCHF] virus, dengue, Ebola virus, Bundibugyo virus, Sudan virus, hantaviruses [Hantaan and Seoul], hepatitis E, Marburg, Nipah virus, o'nyong-nyong virus, Rift Valley fever virus, West Nile virus, and yellow fever virus), 8 bacteria (
Bartonella
spp.,
Brucella
spp.,
Coxiella burnetii
,
Leptospira
spp.,
Rickettsia
spp.,
Salmonella enterica
and
Salmonella enterica
serovar Typhi, and
Yersinia pestis
), and 3 protozoa (
Leishmania
spp.,
Plasmodium
spp., and
Trypanosoma brucei
). Two extrinsic controls (phocine herpesvirus 1 and bacteriophage MS2) were included to ensure extraction and amplification efficiency. Analytical validation was performed on spiked specimens for linearity, intra-assay precision, interassay precision, limit of detection, and specificity. The performance of the card on clinical specimens was evaluated with 1,050 blood samples by comparison to the individual real-time PCR assays, and the TAC exhibited an overall 88% (278/315; 95% confidence interval [CI], 84% to 92%) sensitivity and a 99% (5,261/5,326, 98% to 99%) specificity. This TaqMan array card can be used in field settings as a rapid screen for outbreak investigation or for the surveillance of pathogens, including Ebola virus.
Cytomegalovirus-Specific CD4 T Cells Are Cytolytic and Mediate Vaccine Protection
Authors: Verma Shilpi , Weiskopf Daniela , Gupta Ankan , McDonald Bryan , Peters Bjoern , Sette Alessandro , Benedict Chris A.
Published: October 2015

CD4 T cells provide protection against cytomegalovirus (CMV) and other persistent viruses, and the ability to quantify and characterize epitope-specific responses is essential to gain a more precise understanding of their effector roles in this regard. Here, we report the first two I-A
d
-restricted CD4 T cell responses specific for mouse CMV (MCMV) epitopes and use a major histocompatibility complex class II (MHC-II) tetramer to characterize their phenotypes and functions. We demonstrate that MCMV-specific CD4 T cells can express high levels of granzyme B and kill target cells in an epitope- and organ-specific manner. In addition, CD4 T cell epitope vaccination of immunocompetent mice reduced MCMV replication in the same organs where CD4 cytotoxic T lymphocyte (CTL) activity was observed. Together, our studies show that MCMV epitope-specific CD4 T cells have the potential to mediate antiviral defense by multiple effector mechanisms
in vivo
.

IMPORTANCE
CD4 T cells mediate immune protection by using their T cell receptors to recognize specific portions of viral proteins, called epitopes, that are presented by major histocompatibility complex class II (MHC-II) molecules on the surfaces of professional antigen-presenting cells (APCs). In this study, we discovered the first two epitopes derived from mouse cytomegalovirus (MCMV) that are recognized by CD4 T cells in BALB/c mice, a mouse strain commonly used to study the pathogenesis of this virus infection. Here, we report the sequences of these epitopes, characterize the CD4 T cells that recognize them to fight off MCMV infection, and show that we can use the epitopes to vaccinate mice and protect against MCMV.
Interferon Kappa Inhibits Human Papillomavirus 31 Transcription by Inducing Sp100 Proteins
Authors: Habiger Christina , Jäger Günter , Walter Michael , Iftner Thomas , Stubenrauch Frank
Published: October 2015

High-risk human papillomaviruses (hr-HPV) establish persistent infections in keratinocytes, which can lead to cancer of the anogenital tract. Interferons (IFNs) are a family of secreted cytokines that induce IFN-stimulated genes (ISGs), many of which display antiviral activities. Transcriptome studies have indicated that established hr-HPV-positive cell lines display a reduced expression of ISGs, which correlates with decreased levels of interferon kappa (IFN-κ), a type I IFN constitutively expressed in keratinocytes. Prior studies have also suggested that IFN-β has anti-hr-HPV activity but the underlying mechanisms are not well understood. The downregulation of IFN-κ by hr-HPV raises the possibility that IFN-κ has anti-HPV activity. Using doxycycline-inducible IFN-κ expression in CIN612-9E cells, which maintain extrachromosomally replicating HPV31 genomes, we demonstrated that IFN-κ inhibits the growth of these cells and reduces viral transcription and replication. Interestingly, the initiation of viral early transcription was already inhibited at 4 to 6 h after IFN-κ expression. This was also observed with recombinant IFN-β, suggesting a common mechanism of IFNs. Transcriptome sequencing (RNA-seq) analysis identified 1,367 IFN-κ-regulated genes, of which 221 were modulated >2-fold. The majority of those (71%) matched known ISGs, confirming that IFN-κ acts as abona fidetype I IFN in hr-HPV-positive keratinocytes. RNA interference (RNAi) and cotransfection experiments indicated that the inhibition of viral transcription is mainly due to the induction of Sp100 proteins by IFN-κ. Consistent with published data showing that Sp100 acts as a restriction factor for HPV18 infection, our results suggest that hr-HPV target IFN-κ to prevent Sp100 expression and identify Sp100 as an ISG with anti-HPV activity.

IMPORTANCEHigh-risk HPV can establish persistent infections which may progress to anogenital cancers. hr-HPV interfere with the expression of interferon (IFN)-stimulated genes (ISGs), which is due to reduced levels of IFN-κ, an IFN that is constitutively expressed in human keratinocytes. This study reveals that IFN-κ rapidly inhibits HPV transcription and that this is due to the induction of Sp100 proteins. Thus, Sp100 represents an ISG for hr-HPV.
Targeting Swine Leukocyte Antigen Class I Molecules for Proteasomal Degradation by the nsp1α Replicase Protein of the Chinese Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus Strain JXwn06
Authors: Du Jige , Ge Xinna , Liu Ying , Jiang Ping , Wang Zhe , Zhang Ruimin , Zhou Lei , Guo Xin , Han Jun , Yang Hanchun
Published: October 2015

Porcine reproductive and respiratory syndrome virus (PRRSV) is a critical pathogen of swine, and infections by this virus often result in delayed, low-level induction of cytotoxic T lymphocyte (CTL) responses in pigs. Here, we report that a Chinese highly pathogenic PRRSV strain possessed the ability to downregulate swine leukocyte antigen class I (SLA-I) molecules on the cell surface of porcine alveolar macrophages and target them for degradation in a manner that was dependent on the ubiquitin-proteasome system. Moreover, we found that the nsp1α replicase protein contributed to this property of PRRSV. Further mutagenesis analyses revealed that this function of nsp1α required the intact molecule, including the zinc finger domain, but not the cysteine protease activity. More importantly, we found that nsp1α was able to interact with both chains of SLA-I, a requirement that is commonly needed for many viral proteins to target their cellular substrates for proteasomal degradation. Together, our findings provide critical insights into the mechanisms of how PRRSV might evade cellular immunity and also add a new role for nsp1α in PRRSV infection.

IMPORTANCE
PRRSV infections often result in delayed, low-level induction of CTL responses in pigs. Deregulation of this immunity is thought to prevent the virus from clearance in an efficient and timely manner, contributing to persistent infections in swineherds. Our studies in this report provide critical insight into the mechanism of how PRRSV might evade CTL responses. In addition, our findings add a new role for nsp1α, a critical viral factor involved in antagonizing host innate immunity.
Env-Specific IgA from Viremic HIV-Infected Subjects Compromises Antibody-Dependent Cellular Cytotoxicity
Authors: Ruiz María Julia , Ghiglione Yanina , Falivene Juliana , Laufer Natalia , Holgado María Pía , Socías María Eugenia , Cahn Pedro , Sued Omar , Giavedoni Luis , Salomón Horacio , Gherardi María Magdalena , Rodríguez Ana María , Turk Gabriela
Published: October 2015

Elucidating the factors that modulate HIV-specific antibody-dependent cellular cytotoxicity (ADCC) will help in understanding its role in HIV immunity. The aim of this study was to determine whether IgA could modify the magnitude of ADCC in HIV infection, abrogating its protective role. Plasma samples from 20 HIV-positive (HIV+) subjects enrolled during primary HIV infection (PHI), 10 chronically infected subjects (chronic), and 7 elite controllers (EC) were used. ADCC was determined by using a fluorometric ADCC assay, before and after removal of plasma IgA. Data were analyzed by using nonparametric statistics. ADCC was documented in 80% of PHI enrollment samples and in 100% of PHI 12-month, chronic, and EC samples; it peaked after acute infection, reached a plateau in chronic infection, and decreased after initiation of antiretroviral treatment (ART). Significant associations between ADCC and disease progression were found only after removal of plasma IgA from 12-month PHI samples: the magnitude of ADCC not only increased after IgA removal but also correlated with CD4+T-cell preservation. This work provides evidence that gp120-specific IgA was capable of modifying ADCC responses during natural HIV infection for the first time and adds to similar evidence provided in other settings. Furthermore, it underscores the complexity of the ADCC phenomenon and will help in an understanding of its underlying mechanisms.

IMPORTANCEAlthough the induction of ADCC-mediating antibodies in HIV-infected subjects has been extensively documented, the association of these antibodies with protection from disease progression is poorly understood. Here, we demonstrate that plasma IgA is a factor capable of modifying the magnitude of IgG-mediated ADCC in HIV infection, mitigating its beneficial effect. These results help in understanding why previous studies failed to demonstrate correlations between ADCC and disease progression, and they also contribute to the notion that an HIV vaccine should stimulate the production of ADCC-mediating IgG antibodies but not IgA.
Characterization of a Herpes Simplex Virus 1 (HSV-1) Chimera in Which the Us3 Protein Kinase Gene Is Replaced with the HSV-2 Us3 Gene
Authors: Shindo Keiko , Kato Akihisa , Koyanagi Naoto , Sagara Hiroshi , Arii Jun , Kawaguchi Yasushi
Published: October 2015

Us3 protein kinases encoded by herpes simplex virus 1 (HSV-1) and 2 (HSV-2) play important roles in viral replication and pathogenicity. To investigate type-specific differences between HSV-1 Us3 and HSV-2 Us3 in cells infected by viruses with all the same viral gene products except for their Us3 kinases, we constructed and characterized a recombinant HSV-1 in which its Us3 gene was replaced with the HSV-2 Us3 gene. Replacement of HSV-1 Us3 with HSV-2 Us3 had no apparent effect on viral growth in cell cultures or on the range of proteins phosphorylated by Us3. HSV-2 Us3 efficiently compensated for HSV-1 Us3 functions, including blocking apoptosis, controlling infected cell morphology, and downregulating cell surface expression of viral envelope glycoprotein B. In contrast, replacement of HSV-1 Us3 by HSV-2 Us3 changed the phosphorylation status of UL31 and UL34, which are critical viral regulators of nuclear egress. It also caused aberrant localization of these viral proteins and aberrant accumulation of primary enveloped virions in membranous vesicle structures adjacent to the nuclear membrane, and it reduced viral cell-cell spread in cell cultures and pathogenesis in mice. These results clearly demonstrated biological differences between HSV-1 Us3 and HSV-2 Us3, especially in regulation of viral nuclear egress and phosphorylation of viral regulators critical for this process. Our study also suggested that the regulatory role(s) of HSV-1 Us3, which was not carried out by HSV-2 Us3, was important for HSV-1 cell-cell spread and pathogenesis
in vivo
.

IMPORTANCE
A previous study comparing the phenotypes of HSV-1 and HSV-2 suggested that the HSV-2 Us3 kinase lacked some of the functions of HSV-1 Us3 kinase. The difference between HSV-1 and HSV-2 Us3 kinases appeared to be due to the fact that some Us3 phosphorylation sites in HSV-1 proteins are not conserved in the corresponding HSV-2 proteins. Therefore, we generated recombinant HSV-1 strains YK781 (Us3-chimera) with HSV-2 Us3 and its repaired virus YK783 (Us3-repair) with HSV-1 Us3, to compare the activities of HSV-1 Us3 and HSV-2 Us3 in cells infected by viruses with the same HSV-1 gene products except for their Us3 kinases. We report here that some processes in viral nuclear egress and pathogenesis
in vivo
that have been attributed to HSV-1 Us3 could not be carried out by HSV-2 Us3. Therefore, our study clarified the biological differences between HSV-1 Us3 and HSV-2 Us3, which may be relevant to viral pathogenesis
in vivo
.
Specific Hopanoid Classes Differentially Affect Free-Living and Symbiotic States of
Bradyrhizobium diazoefficiens
Authors: Kulkarni Gargi , Busset Nicolas , Molinaro Antonio , Gargani Daniel , Chaintreuil Clemence , Silipo Alba , Giraud Eric , Newman Dianne K.
Published: October 2015

A better understanding of how bacteria resist stresses encountered during the progression of plant-microbe symbioses will advance our ability to stimulate plant growth. Here, we show that the symbiotic system comprising the nitrogen-fixing bacterium
Bradyrhizobium diazoefficiens
and the legume
Aeschynomene afraspera
requires hopanoid production for optimal fitness
.
While methylated (2Me) hopanoids contribute to growth under plant-cell-like microaerobic and acidic conditions in the free-living state, they are dispensable during symbiosis. In contrast, synthesis of extended (C
35
) hopanoids is required for growth microaerobically and under various stress conditions (high temperature, low pH, high osmolarity, bile salts, oxidative stress, and antimicrobial peptides) in the free-living state and also during symbiosis. These defects might be due to a less rigid membrane resulting from the absence of free or lipidA-bound C
35
hopanoids or the accumulation of the C
30
hopanoid diploptene. Our results also show that C
35
hopanoids are necessary for symbiosis only with the host
Aeschynomene afraspera
but not with soybean. This difference is likely related to the presence of cysteine-rich antimicrobial peptides in
Aeschynomene
nodules that induce drastic modification in bacterial morphology and physiology. The study of hopanoid mutants in plant symbionts thus provides an opportunity to gain insight into host-microbe interactions during later stages of symbiotic progression, as well as the microenvironmental conditions for which hopanoids provide a fitness advantage.

IMPORTANCE
Because bradyrhizobia provide fixed nitrogen to plants, this work has potential agronomical implications. An understanding of how hopanoids facilitate bacterial survival in soils and plant hosts may aid the engineering of more robust agronomic strains, especially relevant in regions that are becoming warmer and saline due to climate change. Moreover, this work has geobiological relevance: hopanes, molecular fossils of hopanoids, are enriched in ancient sedimentary rocks at discrete intervals in Earth history. This is the first study to uncover roles for 2Me- and C
35
hopanoids in the context of an ecological niche that captures many of the stressful environmental conditions thought to be important during (2Me)-hopane deposition. Though much remains to be done to determine whether the conditions present within the plant host are shared with niches of relevance to the rock record, our findings represent an important step toward identifying conserved mechanisms whereby hopanoids contribute to fitness.
Nonreplicating Influenza A Virus Vaccines Confer Broad Protection against Lethal Challenge
Authors: Baz Mariana , Boonnak Kobporn , Paskel Myeisha , Santos Celia , Powell Timothy , Townsend Alain , Subbarao Kanta
Published: October 2015

New vaccine technologies are being investigated for their ability to elicit broadly cross-protective immunity against a range of influenza viruses. We compared the efficacies of two intranasally delivered nonreplicating influenza virus vaccines (H1 and H5 S-FLU) that are based on the suppression of the hemagglutinin signal sequence, with the corresponding H1N1 and H5N1 cold-adapted (
ca
) live attenuated influenza virus vaccines in mice and ferrets. Administration of two doses of H1 or H5 S-FLU vaccines protected mice and ferrets from lethal challenge with homologous, heterologous, and heterosubtypic influenza viruses, and two doses of S-FLU and
ca
vaccines yielded comparable effects. Importantly, when ferrets immunized with one dose of H1 S-FLU or
ca
vaccine were challenged with the homologous H1N1 virus, the challenge virus failed to transmit to naive ferrets by the airborne route. S-FLU technology can be rapidly applied to any emerging influenza virus, and the promising preclinical data support further evaluation in humans.

IMPORTANCE
Influenza viruses continue to represent a global public health threat, and cross-protective vaccines are needed to prevent seasonal and pandemic influenza. Currently licensed influenza vaccines are based on immunity to the hemagglutinin protein that is highly variable. However, T cell responses directed against highly conserved viral proteins contribute to clearance of the virus and confer broadly cross-reactive and protective immune responses against a range of influenza viruses. In this study, two nonreplicating pseudotyped influenza virus vaccines were compared with their corresponding live attenuated influenza virus vaccines, and both elicited robust protection against homologous and heterosubtypic challenge in mice and ferrets, making them promising candidates for further evaluation in humans.
Herpesvirus Entry Mediator on Radiation-Resistant Cell Lineages Promotes Ocular Herpes Simplex Virus 1 Pathogenesis in an Entry-Independent Manner
Authors: Edwards Rebecca G. , Kopp Sarah J. , Karaba Andrew H. , Wilcox Douglas R. , Longnecker Richard
Published: October 2015

Ocular herpes simplex virus 1 (HSV-1) infection leads to a potentially blinding immunoinflammatory syndrome, herpes stromal keratitis (HSK). Herpesvirus entry mediator (HVEM), a widely expressed tumor necrosis factor (TNF) receptor superfamily member with diverse roles in immune signaling, facilitates viral entry through interactions with viral glycoprotein D (gD) and is important for HSV-1 pathogenesis. We subjected mice to corneal infection with an HSV-1 mutant in which HVEM-mediated entry was specifically abolished and found that the HVEM-entry mutant produced clinical disease comparable to that produced by the control virus. HVEM-mediated induction of corneal cytokines, which correlated with an HVEM-dependent increase in levels of corneal immune cell infiltrates, was also gD independent. Given the complexity of HVEM immune signaling, we used hematopoietic chimeric mice to determine which HVEM-expressing cells mediate HSV-1 pathogenesis in the eye. Regardless of whether the donor was a wild-type (WT) or HVEM knockout (KO) strain, HVEM KO recipients were protected from ocular HSV-1, suggesting that HVEM on radiation-resistant cell types, likely resident cells of the cornea, confers wild-type-like susceptibility to disease. Together, these data indicate that HVEM contributes to ocular pathogenesis independently of entry and point to an immunomodulatory role for this protein specifically on radiation-resistant cells.

IMPORTANCEImmune privilege is maintained in the eye in order to protect specialized ocular tissues, such as the translucent cornea, from vision-reducing damage. Ocular herpes simplex virus 1 (HSV-1) infection can disrupt this immune privilege, provoking a host response that ultimately brings about the majority of the damage seen with the immunoinflammatory syndrome herpes stromal keratitis (HSK). Our previous work has shown that HVEM, a host TNF receptor superfamily member that also serves as a viral entry receptor, is a critical component contributing to ocular HSV-1 pathogenesis, although its precise role in this process remains unclear. We hypothesized that HVEM promotes an inflammatory microenvironment in the eye through immunomodulatory actions, enhancing disease after ocular inoculation of HSV-1. Investigating the mechanisms responsible for orchestrating this aberrant immune response shed light on the initiation and maintenance of HSK, one of the leading causes of infectious blindness in the developed world.
Wernicke encephalopathy and pellagra in an alcoholic and malnourished patient
Authors: Terada Norihiko , Kinoshita Kensuke , Taguchi Shijima , Tokuda Yasuharu
Published: October 2015

Deficiency of multiple vitamins can be identified in alcoholic and malnourished patients. We report a patient with Wernicke encephalopathy, a B1 deficiency and pellagra, a niacin deficiency. A 61-year-old Japanese man presented with generalised weakness. He had drunk alcohol heavily for more than a year without eating adequate meals. Physical examination showed disorientation, eye movement impairment, muscle wasting and a rash over the limbs. Multivitamin supplementations improved all the symptoms. Pellagra causes dementia, diarrhoea, or dermatitis, and can mimic non-specific erythaema in alcoholics. The differential diagnosis between pellagra and non-specific erythaema is important because of the treatability of pellagra by niacin supplementation.
Access to essential paediatric surgery in the developing world: a case of imperforate anus with rectovaginal and rectocutaneous fistulas left untreated
Authors: Vinluan Marilyn L , Olveda Remigio M , Ortanez Clive K , Abellera Modesto , Olveda David U , Chy Delia C , Ross Allen G
Published: October 2015

Anorectal malformations consist of a wide spectrum of conditions which can affect both sexes and involve the distal anus and rectum as well as the urinary and genital tracts. Patients have the best chance of a good functional outcome if the condition is diagnosed early and efficient anatomic repair is promptly instituted. This report describes a rare case of imperforate anus associated with both rectovaginal and rectocutaneous fistulas in a 6-year-old Filipino girl. The case highlights shortcomings in the healthcare delivery system combined with socio-economic factors that contributed to the delay in both diagnosis and the institution of adequate treatment. Care and preventive measures that can be implemented in low-resource settings to reduce the impact of birth defects are also discussed.
Diagnostic accuracy of calculated serum osmolarity to predict dehydration in older people: adding value to pathology laboratory reports
Authors: Hooper Lee , Abdelhamid Asmaa , Ali Adam , Bunn Diane K , Jennings Amy , John W Garry , Kerry Susan , Lindner Gregor , Pfortmueller Carmen A , Sjöstrand Fredrik , Walsh Neil P , Fairweather-Tait Susan J , Potter John F , Hunter Paul R , Shepstone Lee
Published: October 2015

Objectives

To assess which osmolarity equation best predicts directly measured serum/plasma osmolality and whether its use could add value to routine blood test results through screening for dehydration in older people.

Design

Diagnostic accuracy study.

Participants

Older people (≥65 years) in 5 cohorts: Dietary Strategies for Healthy Ageing in Europe (NU-AGE, living in the community), Dehydration Recognition In our Elders (DRIE, living in residential care), Fortes (admitted to acute medical care), Sjöstrand (emergency room) or Pfortmueller cohorts (hospitalised with liver cirrhosis).

Reference standard for hydration status

Directly measured serum/plasma osmolality: current dehydration (serum osmolality >300 mOsm/kg), impending/current dehydration (≥295 mOsm/kg).

Index tests

39 osmolarity equations calculated using serum indices from the same blood draw as directly measured osmolality.

Results

Across 5 cohorts 595 older people were included, of whom 19% were dehydrated (directly measured osmolality >300 mOsm/kg). Of 39 osmolarity equations, 5 showed reasonable agreement with directly measured osmolality and 3 had good predictive accuracy in subgroups with diabetes and poor renal function. Two equations were characterised by narrower limits of agreement, low levels of differential bias and good diagnostic accuracy in receiver operating characteristic plots (areas under the curve >0.8). The best equation was osmolarity=1.86×(Na++ K+)+1.15×glucose+urea+14 (all measured in mmol/L). It appeared useful in people aged ≥65 years with and without diabetes, poor renal function, dehydration, in men and women, with a range of ages, health, cognitive and functional status.

Conclusions

Some commonly used osmolarity equations work poorly, and should not be used. Given costs and prevalence of dehydration in older people we suggest use of the best formula by pathology laboratories using a cutpoint of 295 mOsm/L (sensitivity 85%, specificity 59%), to report dehydration risk opportunistically when serum glucose, urea and electrolytes are measured for other reasons in older adults.

Trial registration numbers:

DRIE: Research Register for Social Care, 122273; NU-AGE: ClinicalTrials.govNCT01754012.
The use of patient experience survey data by out-of-hours primary care services: a qualitative interview study
Authors: Barry Heather E , Campbell John L , Asprey Anthea , Richards Suzanne H
Published: October 2015

Background

English National Quality Requirements mandate out-of-hours primary care services to routinely audit patient experience, but do not state how it should be done.

Objectives

We explored how providers collect patient feedback data and use it to inform service provision. We also explored staff views on the utility of out-of-hours questions from the English General Practice Patient Survey (GPPS).

Methods

A qualitative study was conducted with 31 staff (comprising service managers, general practitioners and administrators) from 11 out-of-hours primary care providers in England, UK. Staff responsible for patient experience audits within their service were sampled and data collected via face-to-face semistructured interviews.

Results

Although most providers regularly audited their patients’ experiences by using patient surveys, many participants expressed a strong preference for additional qualitative feedback. Staff provided examples of small changes to service delivery resulting from patient feedback, but service-wide changes were not instigated. Perceptions that patients lacked sufficient understanding of the urgent care system in which out-of-hours primary care services operate were common and a barrier to using feedback to enable change. Participants recognised the value of using patient experience feedback to benchmark services, but perceived weaknesses in the out-of-hours items from the GPPS led them to question the validity of using these data for benchmarking in its current form.

Conclusions

The lack of clarity around how out-of-hours providers should audit patient experience hinders the utility of the National Quality Requirements. Although surveys were common, patient feedback data had only a limited role in service change. Data derived from the GPPS may be used to benchmark service providers, but refinement of the out-of-hours items is needed.
BET 1: is long-term Rivaroxaban superior to Warfarin in pulmonary embolism?
Authors: Morgan Christopher , Body Richard
Published: October 2015

A short cut review was carried out to establish whether Rivaroxaban is superior to warfarin in the management of pulmonary embolism. Three studies were directly relevant to the question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of these papers are tabulated. The clinical bottom line is that Rivaroxaban is a promising alternative to warfarin in the management of pulmonary embolism.