bluuebunny/crossref_metadata_2025 huggingface dataset:
Mobile phone‐based interactive voice response as a tool for improving access to healthcare in remote areas in Ghana – an evaluation of user experiences
Authors: Brinkel J. , May J. , Krumkamp R. , Lamshöft M. , Kreuels B. , Owusu‐Dabo E. , Mohammed A. , Bonacic Marinovic A. , Dako‐Gyeke P. , Krämer A. , Fobil J. N.
Published: March 2017

Objectives

To investigate and determine the factors that enhanced or constituted barriers to the acceptance of an mHealth system which was piloted in Asante‐Akim North District of Ghana to support healthcare of children.

Methods

Four semi‐structured focus group discussions were conducted with a total of 37 mothers. Participants were selected from a study population of mothers who subscribed to a pilot mHealth system which used an interactive voice response (IVR) for its operations. Data were evaluated using qualitative content analysis methods. In addition, a short quantitative questionnaire assessed system's usability (SUS).

Results

Results revealed 10 categories of factors that facilitated user acceptance of the IVR system including quality‐of‐care experience, health education and empowerment of women. The eight categories of factors identified as barriers to user acceptance included the lack of human interaction, lack of update and training on the electronic advices provided and lack of social integration of the system into the community. The usability (SUS median: 79.3; range: 65–97.5) of the system was rated acceptable.

Conclusions

The principles of the tested mHealth system could be of interest during infectious disease outbreaks, such as Ebola or Lassa fever, when there might be a special need for disease‐specific health information within populations.
Deleting the DAG kinase Dgk1 augments yeast vacuole fusion through increased Ypt7 activity and altered membrane fluidity
Authors: Miner Gregory E. , Starr Matthew L. , Hurst Logan R. , Fratti Rutilio A.
Published: March 2017

Diacylglycerol (DAG) is a fusogenic lipid that can be produced through phospholipase C activity on phosphatidylinositol 4,5‐bisphosphate [PI(4,5)P2
], or through phosphatidic acid (PA) phosphatase activity. The fusion of Saccharomyces cerevisiae vacuoles requires DAG, PA and PI(4,5)P2
, and the production of these lipids is thought to provide temporally specific stoichiometries that are critical for each stage of fusion. Furthermore, DAG and PA can be interconverted by the DAG kinase Dgk1 and the PA phosphatase Pah1. Previously we found that
pah1
Δ vacuoles were fragmented, blocked in SNARE priming and showed arrested endosomal maturation. In other pathways the effects of deleting
PAH1
can be compensated for by additionally deleting
DGK1
; however, deleting both genes did not rescue the
pah1
Δ vacuolar defects. Deleting
DGK1
alone caused a marked increase in vacuole fusion that was attributed to elevated DAG levels. This was accompanied by a gain in resistance to the inhibitory effects of PA as well as inhibitors of Ypt7 activity. Together these data show that Dgk1 function can act as a negative regulator of vacuole fusion through the production of PA at the cost of depleting DAG and reducing Ypt7 activity.
Stress‐sensitive arterial hypertension, haemodynamic changes and brain metabolites in hypertensive ISIAH rats: MRI investigation
Authors: Seryapina A. A. , Shevelev O. B. , Moshkin M. P. , Markel A. L. , Akulov A. E.
Published: March 2017

New Findings

What is the central question of this study?

Stress‐sensitive arterial hypertension is considered to be controlled by changes in central and peripheral sympathetic regulating mechanisms, which eventually result in haemodynamic alterations and blood pressure elevation. Therefore, study of the early stages of development of hypertension is of particular interest, because it helps in understanding the aetiology of the disease.

What is the main finding and its importance?

Non‐invasive in vivo investigation in ISIAH rats demonstrated that establishment of sustainable stress‐sensitive hypertension is accompanied by a decrease in prefrontal cortex activity and mobilization of hypothalamic processes, with considerable correlations between haemodynamic parameters and individual metabolite ratios.

The study of early development of arterial hypertension in association with emotional stress is of great importance for better understanding of the aetiology and pathogenesis of the hypertensive disease. Magnetic resonance imaging (MRI) was applied to evaluate the changes in haemodynamics and brain metabolites in 1‐ and 3‐month‐old inherited stress‐induced arterial hypertension (ISIAH) rats (10 male rats) with stress‐sensitive arterial hypertension and in control normotensive Wistar Albino Glaxo (WAG) rats (eight male rats). In the 3‐month‐old ISIAH rats, the age‐dependent increase in blood pressure was associated with increased blood flow through the renal arteries and decreased blood flow in the lower part of the abdominal aorta. The renal vascular resistance in the ISIAH rats decreased during ageing, although at both ages it remained higher than the renal vascular resistance in WAG rats. An integral metabolome portrait demonstrated that development of hypertension in the ISIAH rats was associated with an attenuation of the excitatory and energetic activity in the prefrontal cortex, whereas in the WAG rats the opposite age‐dependent changes were observed. In contrast, in the hypothalamus of 3‐month‐old ISIAH rats, an increase in energetic activity and prevalence of excitatory over inhibitory neurotransmitters was noticed. The blood flow through the main arteries showed a positive correlation with glutamate and glutamine levels in the hypothalamus and a negative correlation with the hypothalamic GABA level. The blood pressure values were positively correlated with hypothalamic choline levels. Thus, the early development of stress‐sensitive hypertension in the ISIAH rats is accompanied by considerable changes both in brain metabolite ratios and in the parameters of blood flow through the main arteries.
Control of Vortex Shedding Using a Screen Attached on the Separation Point of a Circular Cylinder and Its Effect on Drag
Authors: Ozkan Gokturk Memduh , Firat Erhan , Akilli Huseyin
Published: March 2017

The control of flow in the wake of a circular cylinder by an attached permeable plate having various porosity ratios was analyzed experimentally using both particle image velocimetry (PIV) and dye visualization techniques. The force measurements were also done in order to interpret the effect of control method on drag coefficient. The diameter of the cylinder and length to diameter ratio of the plate were kept constant as D = 50 mm and L/D = 1.0, respectively. The porosity ratio, β, which can be defined as the ratio of open surface area to the whole body surface area, was taken as β = 0.4, 0.5, 0.6, 0.7, and 0.8 (permeable plates). The study was performed considering deep water flow conditions with a constant Reynolds number of ReD = 5000 based on the cylinder diameter. Each permeable plate was attached on the separation point and the results were compared with the results of cylinder without permeable plate (plain cylinder) in order to understand the control effect. Both qualitative and quantitative results revealed that the permeable plates of 0.4 ≤ β ≤ 0.6 are effective on controlling the unsteady flow structure downstream of the cylinder, i.e., the vortex formation length was increased, turbulent statistics was reduced and vortex shedding frequency was diminished when the permeable plate attached normal to the cylinder surface from the lower separation point. However, the drag force acting on the cylinder was found to be increased due to the increased cross-sectional area.
Effects of Amorphous Ti–Al–B Nanopowder Additives on Combustion in a Single-Cylinder Diesel Engine
Authors: Fisher Brian T. , Cowart Jim S. , Weismiller Michael R. , Huba Zachary J. , Epshteyn Albert
Published: March 2017

Energetic nanoparticles are promising fuel additives due to their high specific surface area, high energy content, and catalytic capability. Novel amorphous reactive mixed-metal nanopowders (RMNPs) containing Ti, Al, and B, synthesized via a sonochemical reaction, have been developed at the Naval Research Laboratory. These materials have higher energy content than commercial nano-aluminum (nano-Al), making them potentially useful as energy-boosting fuel components. This work examines combustion of RMNPs in a single-cylinder diesel engine (Yanmar L48V). Fuel formulations included up to 4 wt % RMNPs suspended in JP-5, and equivalent nano-Al suspensions for comparison. Although the effects were small, both nano-Al and RMNPs resulted in shorter ignition delays, retarded peak pressure locations, decreased maximum heat release rates, and increased burn durations. A similar but larger engine (Yanmar L100V) was used to examine fuel consumption and emissions for a suspension of 8 wt % RMNPs in JP-5 (and 8 wt % nano-Al for comparison). The engine was operated as a genset under constant load with nominal gross indicated mean effective pressure of 6.5 bar. Unfortunately, the RMNP suspension led to deposits on the injector tip around the orifices, while nano-Al suspensions led to clogging in the fuel reservoir and subsequent engine stall. Nevertheless, fuel consumption rate was 17% lower for the nano-Al suspension compared to baseline JP-5 for the time period prior to stall, which demonstrates the potential value of reactive metal powder additives in boosting volumetric energy density of hydrocarbon fuels.
Voice gender and the segregation of competing talkers: Perceptual learning in cochlear implant simulations
Authors: Sullivan Jessica R. , Assmann Peter F. , Hossain Shaikat , Schafer Erin C.
Published: March 2017

Two experiments explored the role of differences in voice gender in the recognition of speech masked by a competing talker in cochlear implant simulations. Experiment 1 confirmed that listeners with normal hearing receive little benefit from differences in voice gender between a target and masker sentence in four- and eight-channel simulations, consistent with previous findings that cochlear implants deliver an impoverished representation of the cues for voice gender. However, gender differences led to small but significant improvements in word recognition with 16 and 32 channels. Experiment 2 assessed the benefits of perceptual training on the use of voice gender cues in an eight-channel simulation. Listeners were assigned to one of four groups: (1) word recognition training with target and masker differing in gender; (2) word recognition training with same-gender target and masker; (3) gender recognition training; or (4) control with no training. Significant improvements in word recognition were observed from pre- to post-test sessions for all three training groups compared to the control group. These improvements were maintained at the late session (one week following the last training session) for all three groups. There was an overall improvement in masked word recognition performance provided by gender mismatch following training, but the amount of benefit did not differ as a function of the type of training. The training effects observed here are consistent with a form of rapid perceptual learning that contributes to the segregation of competing voices but does not specifically enhance the benefits provided by voice gender cues.
Human genetic and metabolite variation reveals that methylthioadenosine is a prognostic biomarker and an inflammatory regulator in sepsis
Authors: Wang Liuyang , Ko Emily R. , Gilchrist James J. , Pittman Kelly J. , Rautanen Anna , Pirinen Matti , Thompson J. Will , Dubois Laura G. , Langley Raymond J. , Jaslow Sarah L. , Salinas Raul E. , Rouse D. Clayburn , Moseley M. Arthur , Mwarumba Salim , Njuguna Patricia , Mturi Neema ,   Wellcome Trust Case Control Consortium 2,   Kenyan Bacteraemia Study Group, Williams Thomas N. , Scott J. Anthony G. , Hill Adrian V. S. , Woods Christopher W. , Ginsburg Geoffrey S. , Tsalik Ephraim L. , Ko Dennis C.
Published: March 2017

The substrate of the methionine salvage pathway, methylthioadenosine, is a predictor of death in sepsis patients.
Polymyxins: Antibacterial Activity, Susceptibility Testing, and Resistance Mechanisms Encoded by Plasmids or Chromosomes
Authors: Poirel Laurent , Jayol Aurélie , Nordmann Patrice
Published: March 2017

Polymyxins are well-established antibiotics that have recently regained significant interest as a consequence of the increasing incidence of infections due to multidrug-resistant Gram-negative bacteria. Colistin and polymyxin B are being seriously reconsidered as last-resort antibiotics in many areas where multidrug resistance is observed in clinical medicine. In parallel, the heavy use of polymyxins in veterinary medicine is currently being reconsidered due to increased reports of polymyxin-resistant bacteria. Susceptibility testing is challenging with polymyxins, and currently available techniques are presented here. Genotypic and phenotypic methods that provide relevant information for diagnostic laboratories are presented. This review also presents recent works in relation to recently identified mechanisms of polymyxin resistance, including chromosomally encoded resistance traits as well as the recently identified plasmid-encoded polymyxin resistance determinant MCR-1. Epidemiological features summarizing the current knowledge in that field are presented.
Mass Spectrometry Approaches for Identification and Quantitation of Therapeutic Monoclonal Antibodies in the Clinical Laboratory
Authors: Ladwig Paula M. , Barnidge David R. , Willrich Maria A. V.
Published: March 2017

Therapeutic monoclonal antibodies (MAbs) are an important class of drugs used to treat diseases ranging from autoimmune disorders to B cell lymphomas to other rare conditions thought to be untreatable in the past. Many advances have been made in the characterization of immunoglobulins as a result of pharmaceutical companies investing in technologies that allow them to better understand MAbs during the development phase. Mass spectrometry is one of the new advancements utilized extensively by pharma to analyze MAbs and is now beginning to be applied in the clinical laboratory setting. The rise in the use of therapeutic MAbs has opened up new challenges for the development of assays for monitoring this class of drugs. MAbs are larger and more complex than typical small-molecule therapeutic drugs routinely analyzed by mass spectrometry. In addition, they must be quantified in samples that contain endogenous immunoglobulins with nearly identical structures. In contrast to an enzyme-linked immunosorbent assay (ELISA) for quantifying MAbs, mass spectrometry-based assays do not rely on MAb-specific reagents such as recombinant antigens and/or anti-idiotypic antibodies, and time for development is usually shorter. Furthermore, using molecular mass as a measurement tool provides increased specificity since it is a first-order principle unique to each MAb. This enables rapid quantification of MAbs and multiplexing. This review describes how mass spectrometry can become an important tool for clinical chemists and especially immunologists, who are starting to develop assays for MAbs in the clinical laboratory and are considering mass spectrometry as a versatile platform for the task.
Targeting and Insertion of Membrane Proteins
Authors: Kuhn Andreas , Koch Hans-Georg , Dalbey Ross E.
Published: March 2017

The insertion and assembly of proteins into the inner membrane of bacteria are crucial for many cellular processes, including cellular respiration, signal transduction, and ion and pH homeostasis. This process requires efficient membrane targeting and insertion of proteins into the lipid bilayer in their correct orientation and proper conformation. Playing center stage in these events are the targeting components, signal recognition particle (SRP) and the SRP receptor FtsY, as well as the insertion components, the Sec translocon and the YidC insertase. Here, we will discuss new insights provided from the recent high-resolution structures of these proteins. In addition, we will review the mechanism by which a variety of proteins with different topologies are inserted into the inner membrane of Gram-negative bacteria. Finally, we report on the energetics of this process and provide information on how membrane insertion occurs in Gram-positive bacteria and
Archaea
. It should be noted that most of what we know about membrane protein assembly in bacteria is based on studies conducted in
Escherichia coli
.
Laboratory Diagnostics for Histoplasmosis
Authors: Azar Marwan M. , Hage Chadi A.
Published: March 2017

The diagnosis of histoplasmosis is based on a multifaceted approach that includes clinical, radiographic, and laboratory evidence of disease. The gold standards for laboratory diagnosis include demonstration of yeast on pathological examination of tissue and isolation of the mold in the culture of clinical specimens; however, antigen detection has provided a rapid, noninvasive, and highly sensitive method for diagnosis and is a useful marker of treatment response. Molecular methods with improved sensitivity on clinical specimens are being developed but are not yet ready for widespread clinical use. This review synthesizes currently available laboratory diagnostics for histoplasmosis, with an emphasis on complexities of testing and performance in various clinical contexts.
Multicenter Evaluation of the Cepheid Xpert Hepatitis C Virus Viral Load Assay
Authors: McHugh M. P. , Wu A. H. B. , Chevaliez S. , Pawlotsky J. M. , Hallin M. , Templeton K. E.
Published: March 2017

Viral load monitoring for hepatitis C virus (HCV) is necessary to diagnose infection and monitor response to therapy, but the tests involved are currently confined to specialist institutions. There is a need for a fast, accurate assay with limited operator input to enhance the access to viral load monitoring. We evaluated the quantification of HCV RNA in serum and plasma by the Cepheid Xpert HCV Viral Load assay in comparison to the Abbott RealTi
m
e HCV assay. Serum and plasma samples were gathered from HCV-infected individuals at four international sites. These were tested with the Xpert HCV Viral Load assay, and results were compared to quantification by the Abbott RealTi
m
e HCV assay. An external quality assessment panel of eight samples was also tested. In total, 614 samples were analyzed in the study, and the qualitative results agreed on the two platforms for 588 (95.8%) samples. Further analysis of 396 samples quantified by both tests showed strong correlation (correlation coefficient
r
= 0.99) across the quantifiable range, with Bland-Altman plot data showing a mean difference (±1.96 standard deviation) of 0.03 ± 0.44 log
10
IU/ml. In the external quality assessment panel, the Xpert HCV Viral Load assay results (quantified in log
10
IU per milliliter) were within 1 standard deviation of the target value for all but one sample, which was also similarly misquantified by the Abbott RealTime HCV assay. The Xpert HCV Viral Load assay performs well compared to a market-leading HCV viral load test and should be considered for instances where rapid near-to-patient testing is required.
Impact of a Rapid Herpes Simplex Virus PCR Assay on Duration of Acyclovir Therapy
Authors: Van Tam T. , Mongkolrattanothai Kanokporn , Arevalo Melissa , Lustestica Maryann , Dien Bard Jennifer
Published: March 2017

Herpes simplex virus (HSV) infections of the central nervous system (CNS) are associated with significant morbidity and mortality rates in children. This study assessed the impact of a direct HSV (dHSV) PCR assay on the time to result reporting and the duration of acyclovir therapy for children with signs and symptoms of meningitis and encephalitis. A total of 363 patients with HSV PCR results from cerebrospinal fluid (CSF) samples were included in this retrospective analysis, divided into preimplementation and postimplementation groups. For the preimplementation group, CSF testing was performed using a laboratory-developed real-time PCR assay; for the postimplementation group, CSF samples were tested using a direct sample-to-answer assay. All CSF samples were negative for HSV. Over 60% of patients from both groups were prescribed acyclovir. The average HSV PCR test turnaround time for the postimplementation group was reduced by 14.5 h (23.6 h versus 9.1 h;
P
< 0.001). Furthermore, 79 patients (43.6%) in the postimplementation group had dHSV PCR results reported <4 h after specimen collection. The mean time from specimen collection to acyclovir discontinuation was 17.1 h shorter in the postimplementation group (31.1 h versus 14 h;
P
< 0.001). The median duration of acyclovir therapy was also significantly reduced in the postimplementation group (29.2 h versus 14.3 h;
P
= 0.01). Our investigation suggests that implementation of rapid HSV PCR testing can decrease turnaround times and the duration of unnecessary acyclovir therapy.
The Deacetylase SIRT1 Regulates the Replication Properties of Human Papillomavirus 16 E1 and E2
Authors: Das Dipon , Smith Nathan , Wang Xu , Morgan Iain M.
Published: March 2017

Human papillomaviruses (HPV) replicate their genomes in differentiating epithelium using the viral proteins E1 and E2 in association with host proteins. While the roles of E1 and E2 in this process are understood, the host factors involved and how they interact with and regulate E1-E2 are not. Our previous work identified the host replication and repair factor TopBP1 as an E2 partner protein essential for optimal E1-E2 replication and for the viral life cycle. The role of TopBP1 in host DNA replication is regulated by the class III deacetylase SIRT1; activation of the DNA damage response prevents SIRT1 deacetylation of TopBP1, resulting in a switch from DNA replication to repair functions for this protein and cell cycle arrest. Others have demonstrated an essential role for SIRT1 in regulation of the HPV31 life cycle; here, we report that SIRT1 can directly regulate HPV16 E1-E2-mediated DNA replication. SIRT1 is part of the E1-E2 DNA replication complex and is recruited to the viral origin of replication in an E1-E2-dependent manner. CRISPR/Cas9 was used to generate C33a clones with undetectable SIRT1 expression and lack of SIRT1 elevated E1-E2 DNA replication, in part due to increased acetylation and stabilization of the E2 protein in the absence of SIRT1. The results demonstrate that SIRT1 is a member of, and can regulate, the HPV16 replication complex. We discuss the potential role of this protein in the viral life cycle.

IMPORTANCE
HPV are causative agents in a number of human diseases, and currently only the symptoms of these diseases are treated. To identify novel therapeutic approaches for combating these diseases, the viral life cycle must be understood in more detail. This report demonstrates that a cellular enzyme, SIRT1, is part of the HPV16 DNA replication complex and is brought to the viral genome by the viral proteins E1 and E2. Using gene editing technology (CRISPR/Cas9), the SIRT1 gene was removed from cervical cancer cells. The consequence of this was that viral replication was elevated, probably due to a stabilization of the viral replication factor E2. The overall results demonstrate that an enzyme with known inhibitors, SIRT1, plays an important role in controlling how HPV16 makes copies of itself. Targeting this enzyme could be a new therapeutic approach for combating HPV spread and disease.
The Product of the Herpes Simplex Virus 2 UL16 Gene Is Critical for the Egress of Capsids from the Nuclei of Infected Cells
Authors: Gao Jie , Hay Thomas J. M. , Banfield Bruce W.
Published: March 2017

The herpes simplex virus (HSV)
UL16
gene is conserved throughout the
Herpesviridae
and encodes a poorly understood tegument protein. The HSV-1 UL16 protein forms complexes with several viral proteins, including UL11, gE, VP22, and UL21. We previously demonstrated that HSV-2
UL21
was essential for virus propagation due to the failure of DNA-containing capsids (C capsids) to exit the nucleus. We hypothesized that if a UL16/UL21 complex was required for nuclear egress, HSV-2 lacking UL16 would have a phenotype similar to that of HSV-2 lacking UL21. Deletion of HSV-2
UL16
(Δ16) resulted in a 950-fold reduction in virus propagation in mouse L cell fibroblasts and a 200-fold reduction in virus propagation in Vero cells that was fully reversed upon the repair of Δ16 (Δ16R) and partially reversed by infecting UL16-expressing cells with Δ16. The kinetics of viral gene expression in cells infected with Δ16 were indistinguishable from those of cells infected with Δ16R or the parental virus. Additionally, similar numbers of capsids were isolated from the nuclei of cells infected with Δ16 and the parental virus. However, transmission electron microscopy, fluorescence
in situ
hybridization experiments, and fluorescent capsid localization assays all indicated a reduction in the ability of Δ16 C capsids to exit the nucleus of infected cells. Taken together, these data indicate that, like
UL21
,
UL16
is critical for HSV-2 propagation and suggest that the UL16 and UL21 proteins may function together to facilitate the nuclear egress of capsids.

IMPORTANCE
HSV-2 is a highly prevalent sexually transmitted human pathogen that is the main cause of genital herpes infections and is fueling the epidemic transmission of HIV in sub-Saharan Africa. Despite important differences in the pathological features of HSV-1 and HSV-2 infections, HSV-2 is understudied compared to HSV-1. Here we demonstrate that a deletion of the HSV-2
UL16
gene results in a substantial inhibition of virus replication due to a reduction in the ability of DNA-containing capsids to exit the nucleus of infected cells. The phenotype of this
UL16
mutant resembles that of an HSV-2
UL21
mutant described previously by our laboratory. Because UL16 and UL21 interact, these findings suggest that a complex containing both proteins may function together in nuclear egress.
Modulation of Hepatitis C Virus-Specific CD8 Effector T-Cell Function with Antiviral Effect in Infectious Hepatitis C Virus Coculture Model
Authors: Ojiro Keisuke , Qu Xiaowang , Cho Hyosun , Park Jang-June , Vuidepot Annelise , Lissin Nikolai , Molloy Peter E. , Bennett Alan , Jakobsen Bent K. , Kaplan David E. , Riley James L. , Chang Kyong-Mi
Published: March 2017

The antiviral effects of hepatitis C virus (HCV)-specific CD8 T cells have been shown in an HCV replicon system but not in an authentic infectious HCV cell culture (HCVcc) system. Here, we developed tools to examine the antigenicity of HCV-infected HLA-A2-positive Huh7.5 hepatoma cells (Huh7.5A2 cells) in activating HCV-specific CD8 T cells and the downstream antiviral effects. Infectious HCV epitope mutants encoding the well-defined genotype 1a-derived HLA-A2-restricted HCV NS3-1073 or NS5-2594 epitope were generated from a genotype 2a-derived HCV clone (Jc1Gluc2A) by site-directed mutagenesis. CD8 T-cell lines specific for NS3-1073 and NS5-2594 were expanded from HCV-seropositive persons by peptide stimulation
in vitro
or engineered from HCV-seronegative donor T cells by transduction of a lentiviral vector expressing HCV-specific T-cell receptors. HCV-specific CD8 T cells were cocultured with Huh7.5 cells that were pulsed with titrating doses of HCV epitope peptides or infected with HCV epitope mutants. HCV-specific CD8 T-cell activation (CD107a, gamma interferon, macrophage inflammatory protein 1β, tumor necrosis factor alpha) was dependent on the peptide concentrations and the relative percentages of HCV-infected Huh7.5A2 cells. HCV-infected Huh7.5A2 cells activated HCV-specific CD8 T cells at levels comparable to those achieved with 0.1 to 2 μM pulsed peptides, providing a novel estimate of the level at which endogenously processed HCV epitopes are presented on HCV-infected cells. While HCV-specific CD8 T-cell activation with cytolytic and antiviral effects was blunted by PD-L1 expression on HCV-infected Huh7.5A2 cells, resulting in the improved viability of Huh7.5A2 cells, PD-1 blockade reversed this effect, producing enhanced cytolytic elimination of HCV-infected Huh7.5A2 cells. Our findings, obtained using an infectious HCVcc system, show that the HCV-specific CD8 T-cell function is modulated by antigen expression levels, the percentage of HCV-infected cells, and the PD-1/PD-L1 pathways and has antiviral and cytotoxic effects.

IMPORTANCE
We developed several novel molecular and immunological tools to study the interactions among HCV, HCV-infected hepatocytes, and HCV-specific CD8 T cells. Using these tools, we show the level at which HCV-infected hepatoma cells present endogenously processed HCV epitopes to HCV-specific CD8 T cells with antiviral and cytotoxic effects. We also show the marked protective effect of PD-L1 expression on HCV-infected hepatoma cells against HCV-specific CD8 T cells.
Glycolysis, Glutaminolysis, and Fatty Acid Synthesis Are Required for Distinct Stages of Kaposi's Sarcoma-Associated Herpesvirus Lytic Replication
Authors: Sanchez Erica L. , Pulliam Thomas H. , Dimaio Terri A. , Thalhofer Angel B. , Delgado Tracie , Lagunoff Michael
Published: March 2017

Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiologic agent of Kaposi's sarcoma (KS). KSHV infection induces and requires multiple metabolic pathways, including the glycolysis, glutaminolysis, and fatty acid synthesis (FAS) pathways, for the survival of latently infected endothelial cells. To determine the metabolic requirements for productive KSHV infection, we induced lytic replication in the presence of inhibitors of different metabolic pathways. We found that glycolysis, glutaminolysis, and FAS are all required for maximal KSHV virus production and that these pathways appear to participate in virus production at different stages of the viral life cycle. Glycolysis and glutaminolysis, but not FAS, inhibit viral genome replication and, interestingly, are required for different early steps of lytic gene expression. Glycolysis is necessary for early gene transcription, while glutaminolysis is necessary for early gene translation but not transcription. Inhibition of FAS resulted in decreased production of extracellular virions but did not reduce intracellular genome levels or block intracellular virion production. However, in the presence of FAS inhibitors, the intracellular virions are noninfectious, indicating that FAS is required for virion assembly or maturation. KS tumors support both latent and lytic KSHV replication. Previous work has shown that multiple cellular metabolic pathways are required for latency, and we now show that these metabolic pathways are required for efficient lytic replication, providing novel therapeutic avenues for KS tumors.

IMPORTANCE
KSHV is the etiologic agent of Kaposi's sarcoma, the most common tumor of AIDS patients. KS spindle cells, the main tumor cells, all contain KSHV, mostly in the latent state, during which there is limited viral gene expression. However, a percentage of spindle cells support lytic replication and production of virus and these cells are thought to contribute to overall tumor formation. Our previous findings showed that latently infected cells are sensitive to inhibitors of cellular metabolic pathways, including glycolysis, glutaminolysis, and fatty acid synthesis. Here we found that these same inhibitors block the production of infectious virus from lytically infected cells, each at a different stage of viral replication. Therefore, inhibition of specific cellular metabolic pathways can both eliminate latently infected cells and block lytic replication, thereby inhibiting infection of new cells. Inhibition of metabolic pathways provides novel therapeutic approaches for KS tumors.
Identifying opportunities for nature engagement in cancer care practice and design: protocol for four-round modified electronic Delphi
Authors: Blaschke Sarah , O'Callaghan Clare C , Schofield Penelope
Published: March 2017

Introduction

Opportunities to engage with nature have shown relevance in experiences of health and recovery of patients with cancer and are attracting interest in cancer care practice and design. Such healthcare innovations can widen the horizon of possible supportive care solutions but require deliberate and rigorous investigation to ensure responsible action is taken and wastage avoided. This protocol outlines a study designed to solicit knowledge from relevant experts drawn from a range of healthcare practitioners, management representatives, designers and researchers to explore levels of opinion consensus for determining opportunities for, and barriers to, providing helpful nature engagement in cancer care settings.

Methods and analysis

A 4-round modified electronic Delphi methodology will be used to conduct a structured, iterative feedback process for querying and synthesising expert opinion. Round 1 administers an open-ended questionnaire to a panel of selected, relevant experts who will consider the own recommendations of patients with cancer for nature engagement (drawn from a preceding investigation) before contributing salient issues (items) with relevance to the topic. Round 2 circulates anonymised summaries of responses back to the experts who verify and, if they wish, reconsider their own responses. Rounds 3 and 4 determine and rank experts' top 10 items using a 10-point Likert-type scale. Descriptive statistics (median and mean scores) will be calculated to indicate the items' relative importance. Levels of consensus will be explored with consensus defined as 75% agreement.

Ethics and dissemination

Ethics approval for this study was obtained from the Institution's Human Research Ethics Committee (blinded for review). It is anticipated that the results will be published in peer-reviewed journals and presented in a variety of forums.
Effect of using cardiovascular risk scoring in routine risk assessment in primary prevention of cardiovascular disease: protocol for an overview of systematic reviews
Authors: Studziński Krzysztof , Tomasik Tomasz , Krzysztoń Janusz , Jóźwiak Jacek , Windak Adam
Published: March 2017

Introduction

Major clinical practice guidelines recommend assessing risk of cardiovascular disease (CVD) using absolute/global/total CVD risk scores. However, the effectiveness of using them in clinical practice, despite publication of numerous randomised controlled trials (RCTs), is still poorly understood. To summarise and analyse current knowledge in this field, we will carry out an overview of existing systematic reviews (SRs). The objective of this overview will be to assess the effect of using cardiovascular risk scoring in routine risk assessment in primary prevention of CVD compared with standard care.

Methods and analysis

We will include SRs and meta-analyses which take into account RCTs and quasi-RCTs investigating the effect of using cardiovascular risk scoring in routine risk assessment in primary prevention of CVD. SRs will be retrieved from 4 bibliographical databases and reference lists of identified reviews. Additionally, the PROSPERO database will be searched for unpublished, ongoing or recently completed SRs. 2 reviewers will assess the SRs independently for eligibility and bias. The data will be extracted to a special form. Any disagreement will be resolved by discussion. In case of lack of consensus, a third author will arbitrate. The overview of SRs will be reported according to the Preferred Reporting Items for Systematic Review and Meta-Analyses (PRISMA) statement.

Ethics and dissemination

Ethics approval is not required for overview of SRs. We will summarise evidence concerning whether use of the absolute/global/total CVD risk scoring tools in primary prevention of CVD is effective and supported with scientific data or not. If we face unsatisfactory confirmation, we will highlight a need for further research and advice on how to plan such a study. We will submit the results of our study for peer-review publication in a journal indexed in the international bibliographic database of biomedical information.
Reduced muscular fatigue after a 12-week leucine-rich amino acid supplementation combined with moderate training in elderly: a randomised, placebo-controlled, double-blind trial
Authors: Reule Claudia A , Scholz Claudia , Schoen Christiane , Brown Niklas , Siepelmeyer Anne , Alt Wilfried W
Published: March 2017

Background

Age-related muscle loss is characterised by a progressing decrease in muscle mass, strength and function. Besides resistance training and physical activity, appropriate nutrition that is rich in protein, especially branched-chain amino acids, is very important to support training effects and positively influence the protein synthesis to degradation ratio.

Aim

The purpose of this study was to evaluate the effect of a 12-week leucine-rich amino acid supplementation in combination with moderate training.

Methods

Forty-eight healthy subjects exercised for 30 min three times per week and received either a leucine-rich amino acid supplementation or a placebo. Before and after supplementation, volunteers performed an exhaustive eccentric exercise protocol. Maximal concentric strength, muscle soreness, creatine kinase (CK), type II collagen collagenase cleavage neoepitope (C2C), C propeptide of type II procollagen (CP2) and safety assessments were performed before exercise and after 3, 24, 48 and 72 hours.

Results

The supplementation with leucine resulted in reduced loss of strength at 0 and 3 hours after downhill walking compared with the placebo (p=0.0439). The reduction of C2C/CP2 ratio deflection was significantly increased (p=0.038) due to leucine compared with the placebo. The same tendency could be observed for the recovery phase. No significant supplement effects for muscle soreness and CK could be observed.

Conclusion

The principle findings show that leucine-rich amino acid supplementation can counteract the negative effects of eccentric exercise. The treatment resulted in a reduction of exercise-induced strength loss.
Sarilumab improves patient-reported outcomes in rheumatoid arthritis patients with inadequate response/intolerance to tumour necrosis factor inhibitors
Authors: Strand Vibeke , Reaney Matthew , Chen Chieh-I , Proudfoot Clare W J , Guillonneau Sophie , Bauer Deborah , Mangan Erin , Graham Neil M H , van Hoogstraten Hubert , Lin Yong , Pacheco-Tena César , Fleischmann Roy
Published: March 2017

Objective

To evaluate effects of the anti-interleukin-6 receptor monoclonal antibody sarilumab administered with conventional synthetic disease-modifying antirheumatic drugs (csDMARDs) on patient-reported outcomes (PROs) in the TARGET trial in patients with rheumatoid arthritis (RA) with inadequate response or intolerance to tumour necrosis factor inhibitors (TNF-IR).

Methods

546 patients (81.9% female, mean age 52.9 years) were randomised to placebo, sarilumab 150 or 200 mg subcutaneously every 2 weeks + csDMARDs. PROs included patient global assessment (PtGA); pain and morning stiffness visual analogue scales; Health Assessment Questionnaire Disability Index (HAQ-DI); Short Form-36 Health Survey (SF-36); FACIT-Fatigue (FACIT-F); Work Productivity Survey-Rheumatoid Arthritis (WPS-RA) and Rheumatoid Arthritis Impact of Disease (RAID). Changes from baseline at weeks 12 and 24 were analysed using a mixed model for repeated measures; post hoc analyses included percentages of patients reporting improvements ≥ minimum clinically important differences (MCID) and scores ≥ normative values.

Results

Sarilumab + csDMARDs doses resulted in improvements from baseline at week 12 vs placebo + csDMARDs in PtGA, pain, HAQ-DI, SF-36 and FACIT-F that were maintained at week 24. Sarilumab improved morning stiffness and reduced the impact of RA on work, family, social/leisure activities participation (WPS-RA) and on patients' lives (RAID). Percentages of patients reporting improvements ≥MCID and ≥ normative scores were greater with sarilumab than placebo.

Conclusions

In patients with TNF-IR RA, 150 and 200 mg sarilumab + csDMARDs resulted in clinically meaningful patient-reported benefits on pain, fatigue, function, participation and health status at 12 and 24 weeks that exceeded placebo + csDMARDs, and were consistent with the clinical profile previously reported.

Trial registration number

NCT01709578; Results.
Subcutaneous fat necrosis: a term in question for skin alteration in pigs with dimpled appearance
Authors: von Altrock Alexandra , Hewicker‐Trautwein Marion , Wolf Petra , Hoeltig Doris , Hennig‐Pauka Isabel
Published: March 2017

In 1957, a case of dimpled skin alterations in pigs on a farm was described. The dimpled appearance of the skin was assumed to be due to subcutaneous fat necrosis. Presumably based on that report, the term ‘fat necrosis’ was adopted and used in textbooks where it can be found until today. In the presented case, two five‐month‐old pigs with multifocal irregular dimpling and cobblestone‐like skin changes resembling skin alterations termed ‘subcutaneous fat necrosis’ were investigated. Histology showed that the thickness of the subcutaneous adipose tissue of the dimpled skin area was markedly lower than that of the cobblestone‐like area. Biochemical analysis revealed that skin from the cobblestone‐like area had a higher proportion of unsaturated fatty acids than the dimpled skin area. The results suggest that the dimpled skin appearance of the pigs examined in this study is due to a lower thickness of the outer backfat layer.
Dynamical behavior of the Richtmyer–Meshkov instability-induced turbulent mixing under multiple shock interactions
Authors: Wang Tao , Tao Gang , Bai Jingsong , Li Ping , Wang Bing , Du Lei , Xiao Jiaxin
Published: March 2017

The dynamical behavior of Richtmyer–Meshkov instability-induced turbulent mixing under multiple shock interactions is investigated by large-eddy simulation. After the initial shockwave–interface interaction, the transmitted wave reverberates between the accelerated interface and the end-wall of the shock tube to form a process of multiple shock interactions. The turbulent mixing zone grows in a different manner under each of the impingements. After the initial shock, it grows as a power law of time. After the reshock and the impingement of the reflected rarefaction wave, it grows with time as a different negative exponential law. When the impingement of the reflected compression wave completes, it grows approximately in a linear fashion. The statistical quantities in the turbulent mixing zone evolve with time in a similar way under multiple impingements, and after the impingement of the reflected compression wave, they all decay asymptotically. Therefore, the turbulent mixing zone behaves in a statistically self-similar pattern. Even though the impingements of different waves result in different abrupt changes of the characteristic scale parameters of mixing turbulence, as a whole, the characteristic scales present a feature of growth, and the characteristic-scale Reynolds numbers present a feature of decay. The mixing flow is continuously anisotropic, yet the anisotropy weakens gradually. Therefore the development of turbulent mixing presents a trend of isotropy.
Parameters identification for the unsteady helical flows of a generalized Oldroyd-B fluid model based on its numerical solution
Authors: Ma Ying , Jiang Xiaoyun , Yu Bo
Published: March 2017

In this paper, we mainly consider the problem of parameter identification for the unsteady helical flows of a generalized Oldroyd-B fluid between two infinitely long coaxial circular cylinders. The implicit finite difference method is employed to obtain the numerical solution for the direct problem. By means of the Levenberg–Marquardt method, the numerical inversion for identifying the three parameters of the model is implemented simultaneously, that is, the Riemann–Liouville time-fractional derivative α, relaxation time λ, and retardation time λr. To test the validity of the proposed numerical method, varied initial guesses and the observed data, whether or not they involve random error, have been used to verify the reliability of the estimation. This paper provides an efficient method to obtain the estimated value of the unknown parameters for the generalized non-Newtonian fluids model.
Development of a Urinary Micro Particle Detection System Based on Spatial Coordinate Tracking Method
Authors: Zhou Xiaomou , Ma Xiaoping
Published: March 2017

In order to accurately detect various micro particles in patient's urinary samples, a urinary micro particle detection system based on spatial coordinate tracking method was developed. A hierarchical microscopic image analysis method was described and the detailed spatial coordinate tracking algorithm was designed. The architecture and the procedure of proposed system were presented. Moreover, the prototype system was introduced and six patients' urinary samples were analyzed to verify the reliability and simplicity of unattended operation.
Investigating the Effects of Visuospatial Memory Secondary Tasks on LCT Driving Performance
Authors: Wu Ling , Hu Yueqi , Zhu Tong , Liu Haoxue
Published: March 2017

Memory demand is associated with increased mental workload. The objective of the present study was to examine the effects of visuospatial memory secondary tasks on driving performance. Memory tasks for the unknown word-figure pairs and recognition tasks for word-figure pairs at two-level difficulties were employed separately to represent working memory’s process and long-term memory’s process. A simulator study was conducted based on the simulation of the standard environment of Lane change test (LCT). The performance of lane keeping, lane change, and secondary tasks was measured by statistical methods. The comprehensive appraisal model was constructed to quantify total driving performance. The results showed that the mean path deviation, steering angle, and lane excursion times increased, and the proportion of correct lane change decreased, with the perceived workload increasing and the total driving performance decreasing in dual-task driving condition. Compared with the simple working memory group, as the difficulty of tasks increased in difficult working memory group, lane change performance degraded and the perceived workload increased. In contrast to difficult working memory group, the performance of lane keeping and lane change increased, while the perceived workload decreased and the total performance increased by about 50% in difficult recognition group. There were few differences between the simple working memory group and simple recognition group. The difficult working memory group had the lowest total driving performance. The results indicate that as the secondary task’s difficulty increases, driving performance will degrade. Performance improves significantly when the working memory process is converted to the recognition process. This trend is more obvious when the memory task assumes to be more difficult.