bluuebunny/crossref_metadata_2025_split huggingface dataset:
Fluorescence Resonance Energy Transfer-Based Intracellular Assay for the Conformation of Hepatitis C Virus Drug Target NS5A
Authors: Bhattacharya Dipankar , Ansari Israrul H. , Mehle Andrew , Striker Rob 
Published: May 2012

ABSTRACT
Nonstructural protein 5A (NS5A) is essential for hepatitis C virus (HCV) replication and assembly and is a critical drug target. Biochemical data suggest large parts of NS5A are unfolded as an isolated protein, but little is known about its folded state in the cell. We used fluorescence resonance energy transfer (FRET) to probe whether or not different regions of NS5A are in close proximity within the cell. Twenty-three separate reporter constructs were created by inserting one or more fluorophores into different positions throughout the three domains of NS5A. FRET efficiency was maximal when donor and acceptor fluorophores were positioned next to each other but also could be observed when the two fluorophores flanked NS5A domain 1 or domain 3. Informatic and biochemical analysis suggests that large portions of the carboxy terminus of NS5A are in an unfolded and disordered state. Quercetin, a natural product known to disrupt NS5A function in cells, specifically disrupted a conformationally specific domain 3 FRET signal. Intermolecular FRET indicated that the NS5A amino termini, but not other regions, are in close proximity in multimeric complexes. Overall, this assay provides a new window on the intracellular conformation(s) of NS5A and how the conformation changes in response to cellular and viral components of the replication and assembly complex as well as antiviral drugs.
B-Lymphocyte Dysfunction in Chronic HIV-1 Infection Does Not Prevent Cross-Clade Neutralization Breadth
Authors: Boliar Saikat , Murphy Megan K. , Tran T. Cameron , Carnathan Diane G. , Armstrong Wendy S. , Silvestri Guido , Derdeyn Cynthia A. 
Published: May 2012

ABSTRACT
Aberrant expression of regulatory receptors programmed death-1 (PD-1) and B- and T-lymphocyte attenuator (BTLA) is linked with dysregulation and exhaustion of T lymphocytes during chronic human immunodeficiency virus type 1 (HIV-1) infection; however, less is known about whether a similar process impacts B-lymphocyte function during HIV-1 infection. We reasoned that disruption of the peripheral B cell compartment might be associated with decreased neutralizing antibody activity. Expression of markers that indicate dysregulation (BTLA and PD-1), immune activation (CD95), and proliferation (Ki-67) was evaluated in B cells from HIV-1-infected viremic and aviremic subjects and healthy subjects, in conjunction with immunoglobulin production and CD4 T cell count. Viral load and cross-clade neutralizing activity in plasma from viremic subjects were also assessed. Dysregulation of B lymphocytes was indicated by a marked disruption of peripheral B cell subsets, increased levels of PD-1 expression, and decreased levels of BTLA expression in viremic subjects compared to aviremic subjects and healthy controls. PD-1 and BTLA were correlated in a divergent fashion with immune activation, CD4 T cell count, and the total plasma IgG level, a functional correlate of B cell dysfunction. Within viremic subjects, the total IgG level correlated directly with cross-clade neutralizing activity in plasma. The findings demonstrate that even in chronically infected subjects in which B lymphocytes display multiple indications of dysfunction, antibodies that mediate cross-clade neutralization breadth continue to circulate in plasma.
Complete Genome Sequence of the First Canine Circovirus
Authors: Kapoor Amit , Dubovi Edward J. , Henriquez-Rivera Jose Angel , Lipkin W. Ian 
Published: May 2012

ABSTRACT
We found a highly divergent circovirus in serum samples from several dogs. Phylogenetic analysis indicates that canine circovirus genotype 1 (CaCV-1) represents the first circovirus reported in dogs and is genetically most closely related to the only known mammalian circovirus, porcine circovirus. Here we report the complete genome sequence of the CaCV-1 strain NY214, which will help toward understanding the evolutionary and pathogenic characteristics of mammalian circoviruses.
Genome Sequence of the First Human Adenovirus Type 14 Isolated in China
Authors: Zhang Qiwei , Seto Donald , Zhao Suhui , Zhu Li , Zhao Wei , Wan Chengsong 
Published: May 2012

ABSTRACT
Emergent pathogens may be examined rapidly at high resolution on a molecular level using genomics, allowing an understanding of their evolution. China is a unique environment for studying pathogens, having a large, dense, and generally closed population. Human adenovirus type 14 (HAdV-14) was originally identified as an acute respiratory disease (ARD) pathogen in The Netherlands (1955), with a second isolation in England (1957). Since then, few reports of this virus appeared until an ARD pathogen with a similar genome caused multiple outbreaks in the United States (2006 to 2009). This report presents the first genome of HAdV-B14 isolated in China (2010). As China experienced two recent outbreaks of an emergent ARD pathogen, HAdV-B55, containing much of the HAdV-B14 genome, the availability of this HAdV-B14 sequence will facilitate studies of the epidemiology of these pathogens, as well as provide a foundation for studying adenovirus evolution and the genesis of emergent pathogens. These observations may be invaluable in predicting possible recombination between wild-type viruses and adenoviral gene delivery vectors, including adenovirus vaccines.
Complete Genome Sequence of a Dengue Virus Serotype 4 Strain Isolated in Guangdong, China
Authors: Zhao Hui , Yu Xue-Dong , Zhang Xia-Yi , Jiang Tao , Hong Wen-Xin , Yu Man , Hu Feng-Yu , Zhu Shun-Ya , Qin E-De , Deng Yong-Qiang , Qin Cheng-Feng , Zhang Fu-Chun 
Published: May 2012

ABSTRACT
Here we report the first complete genome sequence of a dengue virus serotype 4 genotype II strain, GZ30, isolated in Guangzhou, Guangdong Province, China, in 2010. The sequence information provided herein will help us to understand the molecular epidemiology of dengue virus and predict the risk of severe diseases in mainland China.
Epstein-Barr Virus Protein Kinase BGLF4 Targets the Nucleus through Interaction with Nucleoporins
Authors: Chang Chou-Wei , Lee Chung-Pei , Huang Yu-Hao , Yang Pei-Wen , Wang Jiin-Tarng , Chen Mei-Ru 
Published: May 2012

ABSTRACT

BGLF4 of Epstein-Barr virus (EBV) encodes a serine/threonine protein kinase that phosphorylates multiple viral and cellular substrates to optimize the cellular environment for viral DNA replication and the nuclear egress of viral nucleocapsids. BGLF4 is expressed predominantly in the nucleus at early and late stages of virus replication, while a small portion of BGLF4 is distributed in the cytoplasm at the late stage of virus replication and packaged into the virion. Here, we analyzed systematically the functional domains crucial for nuclear localization of BGLF4 and found that both the N and C termini play important modulating roles. Analysis of amino acid substitution mutants revealed that the C terminus of BGLF4 does not contain a conventional nuclear localization signal (NLS). Additionally, deletion of the C-terminal putative helical regions at amino acids 386 to 393 and 410 to 419 diminished the nuclear translocation of BGLF4, indicating that the secondary structure of the C terminus is important for the localization of BGLF4. The green fluorescent protein-fused wild-type or C-terminal helical regions of BGLF4 associate with phenylalanine/glycine repeat-containing nucleoporins (Nups) in nuclear envelope fractionation. Both coimmunoprecipitation and
in vitro
pull-down assays further demonstrated that BGLF4 binds to Nup62 and Nup153. Remarkably, nuclear import assay with permeabilized HeLa cells demonstrated that BGLF4 translocated into nucleus independent of cytosolic factors. Data presented here suggest that BGLF4 employs a novel mechanism through direct interactions with nucleoporins for its nuclear targeting.
Arenavirus Nucleoproteins Prevent Activation of Nuclear Factor Kappa B
Authors: Rodrigo W. W. Shanaka I. , Ortiz-Riaño Emilio , Pythoud Christelle , Kunz Stefan , de la Torre Juan C. , Martínez-Sobrido Luis 
Published: May 2012

ABSTRACT

Arenaviruses include several causative agents of hemorrhagic fever (HF) disease in humans that are associated with high morbidity and significant mortality. Morbidity and lethality associated with HF arenaviruses are believed to involve the dysregulation of the host innate immune and inflammatory responses that leads to impaired development of protective and efficient immunity. The molecular mechanisms underlying this dysregulation are not completely understood, but it is suggested that viral infection leads to disruption of early host defenses and contributes to arenavirus pathogenesis in humans. We demonstrate in the accompanying paper that the prototype member in the family, lymphocytic choriomeningitis virus (LCMV), disables the host innate defense by interfering with type I interferon (IFN-I) production through inhibition of the interferon regulatory factor 3 (IRF3) activation pathway and that the viral nucleoprotein (NP) alone is responsible for this inhibitory effect (
C. Pythoud, W. W. Rodrigo, G. Pasqual, S. Rothenberger, L. Martínez-Sobrido, J. C. de la Torre, and S. Kunz
, J. Virol. 86:7728–7738, 2012). In this report, we show that LCMV-NP, as well as NPs encoded by representative members of both Old World (OW) and New World (NW) arenaviruses, also inhibits the nuclear translocation and transcriptional activity of the nuclear factor kappa B (NF-κB). Similar to the situation previously reported for IRF3, Tacaribe virus NP (TCRV-NP) does not inhibit NF-κB nuclear translocation and transcriptional activity to levels comparable to those seen with other members in the family. Altogether, our findings demonstrate that arenavirus infection inhibits NF-κB-dependent innate immune and inflammatory responses, possibly playing a key role in the pathogenesis and virulence of arenavirus.
An institution-wide handoff task force to standardise and improve physician handoffs
Authors: Horwitz Leora I , Schuster Kevin M , Thung Stephen F , Hersh David C , Fisher Rosemarie L , Shah Nidhi , Cushing William , Nunes Judy , Silverman David G , Jenq Grace Y 
Published: May 2012

BackgroundTransfers of care have become increasingly frequent and complex with shorter inpatient stays and changes in work hour regulations. Potential hazards exist with transfers. There are few reports of institution-wide efforts to improve handoffs.MethodsAn institution-wide physician handoff task force was developed to proactively address issues surrounding handoffs and to ensure a consistent approach to handoffs across the institution.ResultsThis report discusses the authors' experiences with handoff standardisation, provider utilisation of a new electronic medical record-based handoff tool, and implementation of an educational curriculum; future work in developing hospital-wide policies and procedures for transfers; and the authors' consensus on the best methods for monitoring and evaluation of trainee handoffs.ConclusionThe handoff task force infrastructure has enabled the authors to take an institution-wide approach to improving handoffs. The task force has improved patient care by addressing handoffs systematically and consistently and has helped create new strategies for minimising risk in handoffs.
Is lidocaine Bier's block safe?
Authors: Jakeman Nicola , Kaye Philip , Hayward James , Watson David P , Turner Stacy 
Published: May 2012

ObjectivesTo assess the safety profile of lidocaine Bier's block when compared with that of prilocaine.MethodA retrospective audit of patients undergoing Bier's block using 0.5% lidocaine during a 27-month period (April 2008–June 2010) at the Royal United Hospital Bath emergency department.Results416 patients with sufficient data were included in the study; 360 women and 56 men. The mean patient age was 65 years. Complications were reported in 39 cases; transient hypotension/vasovagal episodes and transient mild bradycardia were most frequent. No patients required any medical intervention. There was no occurrence of anaphylaxis, convulsion, hypotensive episodes requiring medical intervention, collapse or death.ConclusionNo clinically significant morbidity or mortality as a consequence of lidocaine Bier's block was demonstrated in this audit.
O14 Investigating the recent infection testing algorithm (RITA): predictors of recent HIV infection among GUM clinic attendees
Authors: Rayment M , Aghaizu A , Toswill J , Murphy G , Hughes G , Delpech V 
Published: May 2012

BackgroundTesting for recent infection with HIV has been part of routine national surveillance in the UK since 2009. These data can be used to estimate HIV seroincidence in populations. For these estimates to be accurate, HIV testing behaviour must be independent of HIV acquisition risk. This is unlikely to be true, as much testing may be motivated or clinically indicated.AimsTo identify demographic and behavioural differences between individuals diagnosed with recent (<6 months) vs longstanding HIV infection, and to assess the possible level of bias introduced by motivated testing.MethodsRecent Infection Testing Algorithm (RITA) results were linked to Genitourinary Medicine Clinic Activity Dataset attendance records (providing data on attendance and sexual health) for the year preceding the date of RITA test and/or HIV diagnosis. Univariate analyses were performed examining age, sexual orientation, GUM clinic attendances, and STI history, to identify predictors of being diagnosed at early stages of HIV infection.ResultsPreliminary analyses show that among 628 newly diagnosed HIV-positive individuals, 14% (85/628) were diagnosed with recent HIV infection. Being diagnosed with a recent HIV infection was positively associated with younger age, men who have sex with men and having been diagnosed with any bacterial STI in the year preceding the HIV diagnosis (see Abstract O14 table 1). Those visiting a sexual health clinic more than twice in the previous year were also more likely to be diagnosed at early stages of HIV infection.Abstract O14 Table 1HIV infection according to patient demographicsVariable% (n) recent HIV infection% (n) longstanding HIV infectionUnadjusted RR of being diagnosed with recent infection (95% CI)Age (mean 35 years, SD 9.98, range 16–66 years)––0.98 (0.96 to 1.00)MSM (vs heterosexual)*76.5 (65)45.5 (180)1.68 (1.43 to 1.97)Previous STI† Chlamydia trachomatis21.2 (18)14.9 (81)1.42 (0.90 to 2.24) Syphilis8.2 (7)7 (38)1.18 (0.54 to 2.55) Gonorrhoea16.5 (14)10.5 (57)1.57 (0.92 to 2.69) Lymphogranuloma venereum (LGV)1.2 (1)0.9 (5)1.28 (0.15 to 10.8) Any STI‡37.6 (32)24.5 (133)1.54 (1.13 to 2.10)Visits in the year preceding date of HIV diagnosis/RITA test† >2 visits68 (34)54.8 (188)1.24 (1.00 to 1.54)*n=481.†Recorded in the same GUM clinic.‡Includes Chlamydia, syphilis, gonorrhoea, LGV.ConclusionsImportant behavioural and demographic differences exist between individuals diagnosed with recent vs established HIV infections. Such differences must be considered when deriving incidence estimates among key at-risk groups. Further work to examine these trends among all RITA results, in particular the relationship with HIV testing patterns, is ongoing.
O16 Use of online assessment tools by risk taking men
Authors: Ward P , Power L , Nardone A , Hartney T , Delpech V 
Published: May 2012

BackgroundCondom use by gay men has slipped since the 1990s. In late 2011 THT (as part of CHAPS) launched a condom campaign for younger men, promoting a web-based HIV risk assessment tool.MethodsThe Clever Dick/Smart Arse campaign encouraged use of an online risk assessment tool with prize incentives. This gave an instant rating on sexual activity in the past year, with a detailed risk report on request. This covered behaviours affecting HIV risk including PEP, poppers, condoms, HIV disclosure, receptive or insertive modality, monogamy, treatment and viral load.Results7240 risk assessments were undertaken in 10 weeks of whom 1810 requested their full risk assessment (“Sexscore”) and agreed to further contact. There were 101 248 page views by over 17 000 unique website visitors with average visit time of 3.03 min. Preliminary analysis of 4574 respondents shows high levels of risky behaviours. The majority were not in regular clinic contact while a third had never had a sexual health screen. 24% had never tested for HIV while 10% were knowingly HIV+ (of whom 62% had undetectable VL throughout the year). Over half (56%) had five or more sexual partners in the year. 53% of HIV+ men and 51% of other men reported UAI with one or more partners and 59% of all participants discussed serostatus before sex.ConclusionGay men will engage about risk taking if feedback is instant and anonymous, including those never in contact with sexual health services. Men reporting risk behaviours can be targeted for subsequent interventions.
O20 Home sampling through social network websites: can we reduce undiagnosed HIV?
Authors: Elliot E , McCowan A , McCormak S , Rossi M , Van Every T 
Published: May 2012

BackgroundApproximately 1 in 20 men who have sex with men (MSM) in London has undiagnosed HIV. It is a national priority to minimise time spent undiagnosed to reduce morbidity, mortality and onward transmission. Sigma reported that 29% of MSM recruited online in 2008 had never tested. We piloted an offer of home HIV sampling for MSM in partnership with the Gaydar social network website.ObjectivesThe objectives of the pilot were to determine (1) uptake of an online HIV risk self-assessment (2) uptake of offer for home testing (3) the proportion of those completing the assessment that is high risk.MethodsEach day 200 MSM in London were invited by Gaydar to assess their HIV risk. “High risk” was defined as unprotected anal sex (UAI) with HIV + partner or of unknown HIV status. Following assessment but regardless of risk, users were offered a postal HIV salivary sampling kit. Reactive results were confirmed in the clinic. Data were analysed using Google Analytics.Results4500 invites were issued between 7 November 2011 and 11 January 2012. 363 (8%) clicked through to the project website, 321 (88%) of whom completed the HIV risk assessment. Of these, 146 (45%) were defined as “high risk”. 209 clicked through for information on home sampling, 82 (39%) of whom had never HIV tested. 132 test kits were requested, 73 (55%) were returned, 4 of which were new HIV diagnoses (3% of kits sent and 5.5% of samples returned).DiscussionAlthough low, the uptake appears to reach a population “less tested” than the population that took part in the Sigma online survey or typical of GUM clinics. The level of risk identified was similar to a clinic population with almost half reporting UAI. 55% of kits were returned and we report 5.5% or 1:18 new diagnoses. The project is now scaling up to message all 120 000 Gaydar users in Central London over 12 months and aims to provide evidence to inform the potential roll out of online strategies to enhance community based HIV testing in the UK.
O21 Should we opportunistically treat rectalChlamydia trachomatisinfection in men who have sex with men presenting with non-specific urethritis?
Authors: Rayment M , Bull L , Mandalia S , Boag F , Nwokolo N , Jones R 
Published: May 2012

BackgroundInfection of the rectum withChlamydia trachomatis(CT) is highly prevalent among men who have sex with men (MSM) attending our service (c. 8%). Our guidelines advise different antibiotics for the treatment of non-specific urethritis (NSU) (azithromycin 1 g as a single dose) and for rectal CT infection (doxycycline 100 mg twice daily for 7 days). MSM receiving treatment for NSU require recall if they are latterly found to have concurrent rectal CT. Should we therefore use doxycycline first line in MSM presenting with NSU?AimsTo determine the proportion of MSM diagnosed with NSU with rectal CT infection; to identify independent predictors of having rectal CT.MethodsThe clinic database identified sequential MSM diagnosed with NSU over 6 months. Co-variables collected: age, sexual history, HIV status, contact history and the presence of rectal symptoms. Only men accepting rectal sampling were included. Univariate and multivariable logistic regression analyses were undertaken.Results566 MSM presenting with NSU were identified, of whom 130 (23%) were diagnosed with CT infection of at least one anatomical site. Of this group, 99 were included in the analysis. The overall prevalence of rectal CT infection was 38% (38/99). In men reporting a history of unprotected receptive anal intercourse (UPRAI) in the preceding 6 months, the prevalence was 61% (31/51); in those without it was 7.1% (14/48). In MSM reporting rectal symptoms, the prevalence was 90% (9/10); in those without: 33% (29/89). In the multivariable model, the only independent predictors of risk of rectal CT infection were UPRAI and rectal symptoms (see Abstract O21 table 1).Abstract O21 Table 1Rate of rectal chlamydia according to history and rectal symptomsVariableTotal (n=99)Number with rectal CT (%) (n=38)Adjusted RR95% CIWald statistics, p ValueHistory of unprotected receptive anal intercourse (UPRAI)Y5131 (60.8)10.002N487 (14.6)0.25(0.07 to 0.82)Rectal symptomsY109 (90)9.54(1.09 to 83.4)0.041N8929 (32.6)1DiscussionFirst line treatment of NSU in MSM could be modified based on the likelihood of concurrent CT infection. In men reporting UPRAI in the preceding 6 months, or reporting rectal symptoms, it would be reasonable to offer treatment of NSU with doxycycline. In all other patients, we propose continuing to use azithromycin first line, with subsequent recall for further treatment as necessary.
C5 Two cases of acute encephalitis with neurological complications as a seroconversion illness in primary HIV infection (PHI)
Authors: Jain L , Lowe S , Crowe G 
Published: May 2012

BackgroundNeurological complications as the presenting feature of PHI are rare and reported in <10% of cases. We describe two cases of sero-conversion illness with rapidly progressive neurological involvement.Case 1A 43-year-old man presented with diarrhoea, vomiting and rash. On examination he had widespread lymphadenopathy, mouth ulcers and tender hepatomegaly. HIV test was positive (avidity index 50%). 2 weeks later he was readmitted with headache and vomiting. On examination he had left sided weakness and left facial droop. CT head was normal. Cerebro-spinal fluid (CSF) showed an elevated protein 2.99 g/l, WC 48, with and numerous polymorphs visualised. Antibiotics for meningitis were commenced. Serial MRI scans were normal. CD4 count was 640 cell/mm and HIV viral load was 197 000. Antiretrovirals (ARV) were commenced at day 14 and symptoms improved within the next 4 days. 2 months later all weakness and headaches have resolved.Case 2A 47-year-old British man presented with a collapse following 2 days of personality change and confusion and 10 days of fever, headaches, diarrhoea, myalgia, photophobia and rash. On admission he was confused, pyrexial with a Glasgow Coma Score of 13. He had a normal CT head, CSF protein was 1.5 g/l, RC <5, WC<5, with no organisms seen. Subsequent CSF viral PCR was negative. MRI brain was also normal. No evidence of sepsis was identified. An HIV test on admission was positive (avidity index 65%). HIV test 3 months earlier had been negative. CD4 count was 594 (16%) cells/mm3and viral load 462 756 copies/ml. ARVs were commenced on day 7, symptoms improved markedly over the following week. Four weeks post discharge confusion had completely resolved but he reported ongoing mild coordination difficulties.DiscussionNeurological complications of seroconversion are rare but do occur. Early recognition is important through prompt HIV testing and the introduction of ARVs may reduce the incidence of neurological sequelae markedly.