bluuebunny/crossref_metadata_2025_split huggingface dataset:
Regulation and Function of Phosphorylation on VP8, the Major Tegument Protein of Bovine Herpesvirus 1
Authors: Zhang Kuan , Afroz Sharmin , Brownlie Robert , Snider Marlene , van Drunen Littel-van den Hurk Sylvia 
Published: February 2015

ABSTRACT

The major tegument protein of bovine herpesvirus 1 (BoHV-1), VP8, is essential for virus replication in cattle. VP8 is phosphorylated
in vitro
by casein kinase 2 (CK2) and BoHV-1 unique short protein 3 (U
S
3). In this study, VP8 was found to be phosphorylated in both transfected and infected cells but was detected as a nonphosphorylated form in mature virions. This suggests that phosphorylation of VP8 is strictly controlled during different stages of the viral life cycle. The regulation and function of VP8 phosphorylation by U
S
3 and CK2 were further analyzed. An
in vitro
kinase assay, site-directed mutagenesis, and liquid chromatography-mass spectrometry were used to identify the active sites for U
S
3 and CK2. The two kinases phosphorylate VP8 at different sites, resulting in distinct phosphopeptide patterns. S
16
is a primary phosphoreceptor for U
S
3, and it subsequently triggers phosphorylation at S
32
. CK2 has multiple active sites, among which T
107
appears to be the preferred residue. Additionally, CK2 consensus motifs in the N terminus of VP8 are essential for phosphorylation. Based on these results, a nonphosphorylated VP8 mutant was constructed and used for further studies. In transfected cells phosphorylation was not required for nuclear localization of VP8. Phosphorylated VP8 appeared to recruit promyelocytic leukemia (PML) protein and to remodel the distribution of PML in the nucleus; however, PML protein did not show an association with nonphosphorylated VP8. This suggests that VP8 plays a role in resisting PML-related host antiviral defenses by redistributing PML protein and that this function depends on the phosphorylation of VP8.

IMPORTANCE
The progression of VP8 phosphorylation over time and its function in BoHV-1 replication have not been characterized. This study demonstrates that activation of S
16
initiates further phosphorylation at S
32
by U
S
3. Additionally, VP8 is phosphorylated by CK2 at several residues, with T
107
having the highest level of phosphorylation. Evidence for a difference in the phosphorylation status of VP8 in host cells and mature virus is presented for the first time. Phosphorylation was found to be a critical modification, which enables VP8 to attract and to redistribute PML protein in the nucleus. This might promote viral replication through interference with a PML-mediated antiviral defense. This study provides new insights into the regulation of VP8 phosphorylation and suggests a novel, phosphorylation-dependent function for VP8 in the life cycle of BoHV-1, which is important in view of the fact that VP8 is essential for virus replication
in vivo
.
Dimerization of Matrix Protein Is Required for Budding of Respiratory Syncytial Virus
Authors: Förster Andreas , Maertens Goedele N. , Farrell Paul J. , Bajorek Monika 
Published: February 2015

ABSTRACT

Respiratory syncytial virus (RSV) infects epithelial cells of the respiratory tract and is a major cause of bronchiolitis and pneumonia in children and the elderly. The virus assembles and buds through the plasma membrane, forming elongated membrane filaments, but details of how this happens remain obscure. Oligomerization of the matrix protein (M) is a key step in the process of assembly and infectious virus production. In addition, it was suggested to affect the conformation of the fusion protein, the major current target for RSV antivirals, in the mature virus. The structure and assembly of M are thus key parameters in the RSV antiviral development strategy. The structure of RSV M was previously published as a monomer. Other paramyxovirus M proteins have been shown to dimerize, and biochemical data suggest that RSV M also dimerizes. Here, using size exclusion chromatography-multiangle laser light scattering, we show that the protein is dimeric in solution. We also crystallized M in two crystal forms and show that it assembles into equivalent dimers in both lattices. Dimerization interface mutations destabilize the M dimer
in vitro
. To assess the biological relevance of dimerization, we used confocal imaging to show that dimerization interface mutants of M fail to assemble into viral filaments on the plasma membrane. Additionally, budding and release of virus-like particles are prevented in M mutants that fail to form filaments. Importantly, we show that M is biologically active as a dimer and that the switch from M dimers to higher-order oligomers triggers viral filament assembly and virus production.

IMPORTANCE
Human respiratory syncytial virus (RSV) is the most frequent cause of infantile bronchiolitis and pneumonia. The enormous burden of RSV makes it a major unmet target for a vaccine and antiviral drug therapy. Oligomerization of the matrix protein is a key step in the process of assembly and production of infectious virus, but the molecular mechanism of RSV assembly is still poorly understood. Here we show that the RSV matrix protein forms dimers in solution and in crystals; the dimer is essential for formation of higher-order oligomers. Destabilizing the dimer interface resulted in the loss of RSV filament formation and a lack of budding of virus-like particles. Importantly, our findings can potentially lead to new structure-based RSV inhibitors targeting the assembly process.
Monoclonal Antibodies against Extracellular Domains of Claudin-1 Block Hepatitis C Virus Infection in a Mouse Model
Authors: Fukasawa Masayoshi , Nagase Shotaro , Shirasago Yoshitaka , Iida Manami , Yamashita Mayo , Endo Kohki , Yagi Kiyohito , Suzuki Tetsuro , Wakita Takaji , Hanada Kentaro , Kuniyasu Hiroki , Kondoh Masuo 
Published: February 2015

ABSTRACT

Hepatitis C virus (HCV) entry into host cells is a complex process requiring multiple host factors, including claudin-1 (CLDN1). Safe and effective therapeutic entry inhibitors need to be developed. We isolated a human hepatic Huh7.5.1-derived cell mutant that is nonpermissive to HCV, and comparative microarray analysis showed that the mutant was CLDN1 defective. Four hybridomas were obtained, which produced monoclonal antibodies (MAbs) that interacted with the parental Huh7.5.1 cell but not with the CLDN1-defective mutant. All MAbs produced by these hybridomas specifically bound to human CLDN1 with a very high affinity and prevented HCV infection of Huh7.5.1 cells in a dose-dependent manner, without apparent cytotoxicity. Two selected MAbs also inhibited HCV infection of human liver-chimeric mice without significant adverse effects. CLDN1 may be a potential target to prevent HCV infection
in vivo
. Anti-CLDN1 MAbs may hence be promising candidates as novel anti-HCV agents.

IMPORTANCE
Safe and effective therapeutic entry inhibitors against hepatitis C virus (HCV) are very useful for combination therapies with other anti-HCV drugs, such as direct-acting antivirals. In this study, we first showed an effective strategy for developing functional monoclonal antibodies (MAbs) against extracellular domains of a multimembrane-spanning target protein, claudin-1 (CLDN1), by using parental cells expressing the intact target membrane protein and target-defective cells. The established MAbs against CLDN1, which had a very high affinity for intact CLDN1, efficiently inhibited
in vitro
and
in vivo
HCV infections. These anti-CLDN1 MAbs are promising leads for novel entry inhibitors against HCV.
HIV-1-Induced Impairment of Dendritic Cell Cross Talk with γδ T Lymphocytes
Authors: Cardone Marco , Ikeda Kyojiro N. , Varano Barbara , Gessani Sandra , Conti Lucia 
Published: February 2015

ABSTRACTThe interplay between dendritic cells (DC) and γδ T lymphocytes represents a network of paracrine and cell contact interactions important for an integrated immune response to pathogens. HIV-1 infection dramatically affects the number and functions of both cell populations, and DC/γδ T cell cross talk may represent a target of virus-induced immune escape. We investigated whether HIV-exposed DC could deliver aberrant signals to interacting γδ T cells. Here we report that the interaction of human γδ T lymphocytes with HIV-1-exposed autologous monocyte-derived DC, but not direct exposure to the virus, impairs lymphocyte expansion and gamma interferon (IFN-γ) production in response to phosphoantigens. This effect is independent of virus strain and occurred in 55% of the donors analyzed. The donor-dependent variation observed relies on the responsiveness of DC to HIV-1 and is strictly related to the capacity of the virus to suppress the maturation-induced expression of interleukin 12 (IL-12). In fact, γδ T cell response to phosphoantigens is almost completely recovered when this cytokine is exogenously added to the DC/lymphocyte cocultures. Interestingly, we show that γδ T lymphocytes are recruited by HIV-1-exposed DC through a CCR5-mediated mechanism and exert a CCL4-mediated control on virus dissemination within DC and susceptible CD4+T lymphocytes. These results demonstrate an association between HIV-induced DC dysfunction and alterations of γδ T cell responses. The aberrant cross talk between these two cell populations may contribute to the pathogenesis of HIV infection by further reducing the strength of antiviral immune response.IMPORTANCEThis study provides new evidence on the mechanisms exploited by HIV-1 to evade the host immune response. We report that HIV-1 impairs the cross talk between DC and γδ T lymphocytes, by reducing the capacity of DC to promote functional γδ T cell activation. Interestingly, the virus does notper seinterfere with γδ T cell activation, thus highlighting the key role of early DC–HIV-1 interaction in this phenomenon. Furthermore, the results obtained unravel the novel role of γδ T cells in controlling HIV-1 dissemination within the DC population as well as virus transfer to susceptible CD4+T lymphocytes. The interactions of DC with innate lymphocytes represent a major control mechanism for an integrated immune response to infection. Understanding how HIV-1 harnesses these pathways may provide important insights on the pathogenesis of disease and offer new opportunities for therapeutic interventions.
Diagnostic accuracy of ultrasonography, MRI and MR arthrography in the characterisation of rotator cuff disorders: a systematic review and meta-analysis
Authors: Roy Jean-Sébastien , Braën Caroline , Leblond Jean , Desmeules François , Dionne Clermont E , MacDermid Joy C , Bureau Nathalie J , Frémont Pierre 
Published: February 2015

BackgroundDifferent diagnostic imaging modalities, such as ultrasonography (US), MRI, MR arthrography (MRA) are commonly used for the characterisation of rotator cuff (RC) disorders. Since the most recent systematic reviews on medical imaging, multiple diagnostic studies have been published, most using more advanced technological characteristics. The first objective was to perform a meta-analysis on the diagnostic accuracy of medical imaging for characterisation of RC disorders. Since US is used at the point of care in environments such as sports medicine, a secondary analysis assessed accuracy by radiologists and non-radiologists.MethodsA systematic search in three databases was conducted. Two raters performed data extraction and evaluation of risk of bias independently, and agreement was achieved by consensus. Hierarchical summary receiver-operating characteristic package was used to calculate pooled estimates of included diagnostic studies.ResultsDiagnostic accuracy of US, MRI and MRA in the characterisation of full-thickness RC tears was high with overall estimates of sensitivity and specificity over 0.90. As for partial RC tears and tendinopathy, overall estimates of specificity were also high (>0.90), while sensitivity was lower (0.67–0.83). Diagnostic accuracy of US was similar whether a trained radiologist, sonographer or orthopaedist performed it.ConclusionsOur results show the diagnostic accuracy of US, MRI and MRA in the characterisation of full-thickness RC tears. Since full thickness tear constitutes a key consideration for surgical repair, this is an important characteristic when selecting an imaging modality for RC disorder. When considering accuracy, cost, and safety, US is the best option.
Incidence and fatality of serious suicide attempts in a predominantly rural population in Shandong, China: a public health surveillance study
Authors: Sun Jiandong , Guo Xiaolei , Zhang Jiyu , Wang Mei , Jia Cunxian , Xu Aiqiang 
Published: February 2015

ObjectivesTo estimate the incidence of serious suicide attempts (SSAs, defined as suicide attempts resulting in either death or hospitalisation) and to examine factors associated with fatality among these attempters.DesignA surveillance study of incidence and mortality. Linked data from two public health surveillance systems were analysed.SettingThree selected counties in Shandong, China.ParticipantsAll residents in the three selected counties.Outcome measuresIncidence rate (per 100 000 person-years) and case fatality rate (%).MethodsRecords of suicide deaths and hospitalisations that occurred among residents in selected counties during 2009–2011 (5 623 323 person-years) were extracted from electronic databases of the Disease Surveillance Points (DSP) system and the Injury Surveillance System (ISS) and were linked by name, sex, residence and time of suicide attempt. A multiple logistic regression model was developed to examine the factors associated with a higher or lower fatality rate.ResultsThe incidence of SSAs was estimated to be 46 (95% CI 44 to 48) per 100 000 person-years, which was 1.5 times higher in rural versus urban areas, slightly higher among females, and increased with age. Among all SSAs, 51% were hospitalised and survived, 9% were hospitalised but later died and 40% died with no hospitalisation. Most suicide deaths (81%) were not hospitalised and most hospitalised SSAs (85%) survived. The fatality rate was 49% overall, but was significantly higher among attempters living in rural areas, who were male, older, with lower education or with a farming occupation. With regard to the method of suicide, fatality was lowest for non-pesticide poisons (7%) and highest for hanging (97%).ConclusionsThe incidence of serious suicide attempts is substantially higher in rural areas than in urban areas of China. The risk of death is influenced by the attempter's sex, age, education level, occupation, method used and season of year.
A tool to improve communication in the neonatal unit
Authors: van de Vijver Margot , Evans Mari 
Published: February 2015

AbstractFrequent and sensitive communication from neonatal staff is important to alleviate parental stress and to ensure that parents understand their baby's condition and progress. It also consequently empowers and involves parents in their baby's care. A lack of regular and informative communication from neonatal staff is a common reason for parental complaint. We sought to assess whether the implementation of a baby diary used as a communication tool would improve parent-staff communication and optimise the parental experience of neonatal care.In this study, parents and carers of babies on our neonatal unit were invited to complete a questionnaire to assess their level of satisfaction with communication by neonatal staff before and after the implementation of the diaries. The diary was designed to act as an aid to improve communication to parents about their baby and thereby complementing face-to face communication and encouraging parental involvement.The study design was a retrospective parental satisfaction questionnaire based on the Department of Health [1] and the National Institute for Health and Care Excellence (NICE) [2] quality standards for specialist neonatal care on communication and parental involvement (n=44, response rate 57%). Following implementation of the diary, the questionnaire was repeated on two occasions: one month post implementation (n=17, response rate 23%) and 15 months post implementation (n=44, response rate 65%).More parents in the post-intervention cohort felt they were receiving regular communication from staff, that their questions and concerns were being addressed, and felt involved in their baby's care than in the pre-intervention cohort. In addition, 100% of parents said they liked reading the diary and 94% felt it added to their understanding of how their baby was doing.Our project demonstrates an improvement in parents' satisfaction with communication from neonatal staff following the implementation of individual baby diaries. The diary is a simple, practical, and cost-effective tool to enhance communication between parents and staff in a neonatal unit.
Out of hours intravenous fluid therapy: a prompt to guide prescribing
Authors: Fehmi Janev , Carpenter Alexander , Townsend Matthew , Sheppeard Rhian , Gannon Elizabeth , Hewitson Lynsey , Auerbach Natalie , Thomas Katherine 
Published: February 2015

AbstractRecent NICE guidance has highlighted the importance of appropriate and safe intravenous fluid use. We aimed to improve the quality of out of hours fluid prescription in a Bristol hospital by ensuring that indications and cautions for fluid therapy were clearly documented at the time of initiation.Time-pressured on-call doctors need quick access to information regarding patients’ care. A documented “fluid plan” allows doctors to undertake a more informed assessment of the patient's fluid balance, leading to safer prescriptions.Our ideal was for 100% of out of hours intravenous fluid prescriptions to be appropriate. Our process measures included the proportion of patients on intravenous fluids who had a documented fluid plan in the medical notes or on the prescription chart on Friday, prior to the weekend on call period. This was defined as mention of indications and/or cautions to fluid therapy.The introduction of a sticker to prompt fluid plan documentation did marginally improve use of fluid plans. It was notable that 96% of these were followed where plans were documented (n=23). Initiation of IV fluid with an accompanying plan is likely to make subsequent fluid prescriptions safer.Rapid turnover of staff and stationary proved significant barriers to consistent implementation of the sticker. Despite these challenges we demonstrated a “proof of concept”, suggesting system modification to include fluid plans is safe and effective.
The endoscopy safety checklist: A longitudinal study of factors affecting compliance in a tertiary referral centre within the United Kingdom
Authors: Matharoo Manmeet , Sevdalis Nick , Thillai Muhunthan , Bouri Sonia , Marjot Thomas , Haycock Adam , Thomas-Gibson Siwan 
Published: February 2015

AbstractGastrointestinal endoscopy is a widely used diagnostic and therapeutic procedure both within the United Kingdom and worldwide. With an increasingly older population the potential for complications is increased. The Wolfson Unit for Endoscopy at St. Mark’s Hospital in London is a tertiary referral centre, which conducts over 14,000 endoscopic procedures annually. However, despite this high throughput, our baseline observations were that the procedure for safety checks was highly variable.Over a seven-day period we conducted a questionnaire-based survey to all staff members involved with endoscopy within our unit. We found that there was little consensus between team members, both in terms of essential safety checks and designating responsibility for the checks. A panel of experts was convened in order to devise a safety checklist and a strategy for increasing compliance with the checklist among all staff members. Using a combination of electronic and physical reminders and incentives, we found that there was a significant increase in completed checklist (53% to 66%, p = 0.021) and decrease in the number of checklists left blank post intervention (10% to 2%, p=0.03). We believe that post implementation validation of safety checklists is an important method to ensure their proper use.
Medication Reconciliation Improvement Through the Use of Video
Authors: Dewan Maya , Kraus Blair , Davis Daniela , McCloskey John 
Published: February 2015

AbstractAccurate medication reconciliation at the time of hospital admission is vital to preventing adverse drug events. Compliance with medication reconciliation in our pediatric intensive care unit was low initially with overall medication reconciliation at 70%. Due to the high front line provider turnover in our unit, we focused on technological reminders for completion and used unique and innovative ways to motivate our supervising staff. Our goal was to reach >95% completion within 24 hours for medication reconciliation for all patients admitted to the pediatric intensive care unit.Pre-pilot discussions focused on examples of errors of medication reconciliation within our own institution resulting in patient harm via traditional power point presentation. The initial pilot phase instituted a job aid on how to add the medication reconciliation completion reminder column. Email updates on completion status began one week after initiation. During the implementation and spread phase, fun interactive videos were used to acknowledge roll out to the full unit. Compliance was monitored and humorous biweekly video updates emphasizing accountability were introduced. In the sustain phase, monthly video updates served as education and a reminder for provider staff.The use of a medication reconciliation completion column and reminder emails resulted in goal completion (>95%) by three weeks post intervention for the pilot unit. Overall medication reconciliation completion also reached goal of >95% completion within the entire unit after three weeks of implementation and spread. Compliance fell below goal so video updates were modified utilizing humor and emphasizing accountability. This resulted in sustained compliance now more than forty weeks post implementation.Unit compliance currently remains >95% completion within 24 hours now more than forty weeks post intervention. The use of the medication reconciliation reminder column resulted in improvement in compliance; however, that improvement was not sustained. The addition of humorous videos highlighting accountability allowed for sustained improvements.
Derivation of the time dependent Gross–Pitaevskii equation with external fields
Authors: Pickl Peter 
Published: February 2015

Using a new method [19], it is possible to derive mean field equations from the microscopic N body Schrödinger evolution of interacting particles without using BBGKY hierarchies. This method also allows for error estimates and can be generalized to systems with external fields, of which both points are relevant from a physics perspective. Recently, this method was used to derive the Hartree equation for singular interactions [11] and the Gross–Pitaevskii equation without positivity condition on the interaction [17] where one had to restrict the scaling behavior of the interaction. Assuming positivity of the interaction, this paper deals with more general scalings including the so-called Gross–Pitaevskii scaling.