bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of binding domains on red blood cell glycophorins for Babesia divergens
Authors: Cursino‐Santos Jeny R. , Halverson Greg , Rodriguez Marilis , Narla Mohandas , Lobo Cheryl A. 
Published: August 2013

BackgroundInvasion of red blood cells (RBCs) is one of the critical points in the lifecycle of Babesia. The parasite does not invade other host cells. Earlier work has shown that GPA and GPB function as putative receptors during parasite invasion. The primary focus of this study was the delineation of parasite‐binding domains on GPA and GPB.Study Design and MethodsThe assay of choice to validate molecules that participate in invasion is an inhibition of invasion assay, in which changes in parasitemia are assessed relative to a wild‐type assay (no inhibitors). Inhibition of invasion can be achieved by modification of different components of the assay or by the addition of competitors of the molecules that participate in invasion. In this study purified antibody fragments to various domains on GPA and GPB were tested for magnitude of inhibition of parasite invasion. Effects on invasion were monitored by assessment of Giemsa‐stained smears every 24 hours.ResultsAmong 10 selected antibodies directed at various epitopes on GPA and GPB, antibodies directed against GPAM epitopes had the most severe effect (up to 35%) on inhibition of invasion, followed by antibodies directed against GPBS epitope (up to 24%).ConclusionThis study confirms the role of RBC glycophorins A and B in Babesia divergens invasion and shows that the GPAM and GPBS epitopes are likely to play an important role in the entry process.
Evaluation of the Recombinant Protein TpF1 of Treponema pallidum for Serodiagnosis of Syphilis
Authors: Jiang Chuanhao , Zhao Feijun , Xiao Jinhong , Zeng Tiebing , Yu Jian , Ma Xiaohua , Wu Haiying , Wu Yimou 
Published: August 2013

ABSTRACTSyphilis is a chronic infection caused byTreponema pallidumsubsp.pallidum, and diagnosis with sensitive and specific methods is a challenging process that is important for its prevention and treatment. In the present study, we established a recombinant protein TpF1-based indirect immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) and a Western blot assay for human and rabbit sera. The 20-kDa recombinant protein TpF1 was detected by Western blotting performed with sera from rabbits immunized with recombinant TpF1 and infected with theT. pallidumNichols strain andT. pallidumclinical isolates but was not detected by Western blotting with sera from uninfected rabbits. The sensitivity of the recombinant protein was determined by screening sera from individuals with primary, secondary, latent, and congenital syphilis (n= 82). The specificity of the recombinant protein was determined by screening sera from uninfected controls (n= 30) and individuals with potentially cross-reactive infections, including Lyme disease (n= 30) and leptospirosis (n= 5). The sensitivities of TpF1-based ELISAs were 93.3%, 100%, 100%, and 100% for primary, secondary, latent, and congenital syphilis, respectively, and the specificities were all 100% for sera from uninfected controls and individuals with potentially cross-reactive infections. In Western blot assays, the sensitivities and specificities of TpF1 for human sera were all 100%. The reactivities of TpF1 with syphilitic sera were proportional to the titers of theT. pallidumparticle agglutination (TPPA) assay. These data indicate that the recombinant protein TpF1 is a highly immunogenic protein in human and rabbit infections and a promising marker for the screening of syphilis.
Deep-Sequencing Analysis of the Association between the Quasispecies Nature of the Hepatitis C Virus Core Region and Disease Progression
Authors: Miura Mika , Maekawa Shinya , Takano Shinichi , Komatsu Nobutoshi , Tatsumi Akihisa , Asakawa Yukiko , Shindo Kuniaki , Amemiya Fumitake , Nakayama Yasuhiro , Inoue Taisuke , Sakamoto Minoru , Yamashita Atsuya , Moriishi Kohji , Enomoto Nobuyuki 
Published: August 2013

ABSTRACT
Variation of core amino acid (aa) 70 of hepatitis C virus (HCV) has been shown recently to be closely correlated with liver disease progression, suggesting that the core region might be present as a quasispecies during persistent infection and that this quasispecies nature might have an influence on the progression of disease. In our investigation, the subjects were 79 patients infected with HCV genotype 1b (25 with chronic hepatitis [CH], 29 with liver cirrhosis [LC], and 25 with hepatocellular carcinoma [HCC]). Deep sequencing of the HCV core region was carried out on their sera by using a Roche 454 GS Junior pyrosequencer. Based on a plasmid containing a cloned HCV sequence (pCV-J4L6S), the background error rate associated with pyrosequencing, including the PCR procedure, was calculated as 0.092 ± 0.005/base. Deep sequencing of the core region in the clinical samples showed a mixture of “mutant-type” Q/H and “wild-type” R at the core aa 70 position in most cases (71/79 [89.9%]), and the ratio of mutant residues to R in the mixture increased as liver disease advanced to LC and HCC. Meanwhile, phylogenetic analysis of the almost-complete core region revealed that the HCV isolates differed genetically depending on the mutation status at core aa 70. We conclude that the core aa 70 mixture ratio, determined by deep sequencing, reflected the status of liver disease, demonstrating a significant association between core aa 70 and disease progression in CH patients infected with HCV genotype 1b.
Dual Split Protein-Based Fusion Assay Reveals that Mutations to Herpes Simplex Virus (HSV) Glycoprotein gB Alter the Kinetics of Cell-Cell Fusion Induced by HSV Entry Glycoproteins
Authors: Atanasiu Doina , Saw Wan Ting , Gallagher John R. , Hannah Brian P. , Matsuda Zene , Whitbeck J. Charles , Cohen Gary H. , Eisenberg Roselyn J. 
Published: August 2013

ABSTRACT

Herpes simplex virus (HSV) entry and cell-cell fusion require glycoproteins gD, gH/gL, and gB. We propose that receptor-activated changes to gD cause it to activate gH/gL, which then triggers gB into an active form. We employed a dual split-protein (DSP) assay to monitor the kinetics of HSV glycoprotein-induced cell-cell fusion. This assay measures content mixing between two cells, i.e., fusion, within the same cell population in real time (minutes to hours). Titration experiments suggest that both gD and gH/gL act in a catalytic fashion to trigger gB. In fact, fusion rates are governed by the amount of gB on the cell surface. We then used the DSP assay to focus on mutants in two functional regions (FRs) of gB, FR1 and FR3. FR1 contains the fusion loops (FL1 and FL2), and FR3 encompasses the crown at the trimer top. All FL mutants initiated fusion very slowly, if at all. However, the fusion rates caused by some FL2 mutants increased over time, so that total fusion by 8 h looked much like that of the WT. Two distinct kinetic patterns, “slow and fast,” emerged for mutants in the crown of gB (FR3), again showing differences in initiation and ongoing fusion. Of note are the fusion kinetics of the gB
syn
mutant (LL871/872AA). Although this mutant was originally included as an ongoing high-rate-of-fusion control, its initiation of fusion is so rapid that it appears to be on a “hair trigger.” Thus, the DSP assay affords a unique way to examine the dynamics of HSV glycoprotein-induced cell fusion.
Efficiency of E2-p7 Processing Modulates Production of Infectious Hepatitis C Virus
Authors: Shanmugam Saravanabalaji , Yi MinKyung 
Published: August 2013

ABSTRACT
Previous studies indicate that the processing of hepatitis C virus (HCV) E2-p7-NS2 precursor mediated by host signal peptidase is relatively inefficient, resulting in the accumulation of E2-p7-NS2 and E2-p7 precursors in addition to E2 in mammalian cells. In this study, we discovered that a significant inhibition of the processing at an E2-p7 junction site is detrimental for HCV production, whether it was caused by the mutations in p7 or by the strategic introduction of a mutation at a terminal residue of E2 to block the signal peptidase-mediated cleavage of this junction site. However, complete separation of E2 and p7 by inserting an encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES) between these two proteins also moderately inhibited virus production. These results indicate that optimal processing of the E2-p7 junction site is critical for efficient HCV production. We further demonstrated that disrupting E2-p7 processing inhibits both NS2 localization to the putative virus assembly sites near lipid droplets (LD) and NS2 interaction with NS3 and E2. However, the impact, if any, of the p7-NS2 processing efficiency on HCV production seems relatively minor. In conclusion, these results imply that effective release of E2 and p7 from the precursor E2-p7 promotes HCV production by enhancing NS2-associated virus assembly complex formation near LD.
Human Rhinovirus Subviral A Particle Binds to Lipid Membranes over a Twofold Axis of Icosahedral Symmetry
Authors: Kumar Mohit , Blaas Dieter 
Published: August 2013

ABSTRACT

Minor group human rhinoviruses bind low-density lipoprotein (LDL) receptors for endocytosis. Once they are inside endosomes, the acidic pH triggers their dissociation from the receptors and conversion into hydrophobic subviral A particles; these attach to the membrane and transfer their single-strand, positive-sense RNA genome into the cytosol. Here, we allowed human rhinovirus 2 (HRV2) A particles, produced
in vitro
by incubation at pH 5.4, to attach to liposomes; cryo-electron microscopy 3-dimensional single-particle image reconstruction revealed that they bind to the membrane around a 2-fold icosahedral symmetry axis.
Can resistance training change the strength, body composition and self-concept of overweight and obese adolescent males? A randomised controlled trial
Authors: Schranz Natasha , Tomkinson Grant , Parletta Natalie , Petkov John , Olds Tim 
Published: August 2013

Background/aimResistance training is an exercise modality at which overweight and obese adolescents can excel and which can therefore positively affect their psychological well-being. The aim of this study was to determine the effect of a 6-month resistance training intervention on the self-concept strength and body composition of overweight and obese adolescent males.Methods56 overweight and obese males aged 13–17 years were randomly allocated to an Intervention (n=30) or Control (n=26) group. Primary (psychological) and secondary (strength and body composition) outcomes were assessed at baseline as well as at 3 (halfway through the intervention), 6 (immediately postintervention) and 12 months follow-up. Random effects mixed modelling was used to determine the effects of the intervention.ResultsStatistically significant differences between the Intervention and Control groups were observed at 3-month and 6-month assessments for exercise self-efficacy, resistance training confidence and self-esteem. Large increases in strength for the Intervention group, relative to Controls, were also observed with no substantial changes in body composition shown for either group. Values for all variables returned to baseline following completion of the programme.ConclusionsA 6-month resistance training intervention can positively affect the self-concept and strength of overweight and obese adolescent boys.
Antiviral therapy for prevention of hepatocellular carcinoma and mortality in chronic hepatitis B: systematic review and meta-analysis
Authors: Thiele Maja , Gluud Lise L , Dahl Emilie K , Krag Aleksander 
Published: August 2013

ObjectivesThe effect of antiviral therapy on clinical outcomes in chronic hepatitis B virus (HBV) is not established. We aimed to assess the effects of interferon and/or nucleos(t)ide analogues versus placebo or no intervention on prevention of hepatocellular carcinoma (HCC) and mortality in chronic HBV.DesignRandom-effects pairwise meta-analysis of randomised trials and observational studies.SettingElectronic and manual searches were combined. Randomised controlled trials (RCTs) were included in the primary analyses. Observational studies were included in sensitivity analyses.Primary and secondary outcome measuresThe primary outcome measures were HCC incidence and mortality. The secondary outcome measure was HCC mortality.ResultsWe included 8 RCTs, 8 prospective cohort studies and 19 case–control studies with a total of 3433 patients allocated to antiviral therapy and 4625 controls. The maximum duration of follow-up was 23 years. Randomised trials found no effect of antiviral therapy on HCC or mortality. Cohort studies found that antiviral therapy increased the risk of HCC (risk ratio 1.43; 95% CI 1.06 to 1.95), whereas case–control studies found a decreased risk of HCC in the intervention group (risk ratio 0.69; 95% CI 0.54 to 0.88). There was a clear difference between the results of RCTs and observational studies (test for subgroup differences, p<0.001). Antiviral therapy did not affect mortality in cohort studies, but reduced mortality in case–control studies (relative risk 0.71; 95% CI 0.54 to 0.93; test for subgroup differences, p=0.406).ConclusionsThe effect of antiviral therapy on clinical outcomes in HBV remains to be established. Although there was a positive effect in the sensitivity analyses, the strength of the evidence does not allow for extrapolation to clinical practice as research design plays an essential role in the overall assessment.Trial registration numberProspero number CRD42013003881.
070 Strategies for Health System Implementation of Guidelines on Overweight and Obesity
Authors: Abrahamian Y , Watson H 
Published: August 2013

BackgroundEvidence-based clinical practice guidelines (CPGs) for adult obesity and overweight provide recommendations to clinicians on interventions for weight loss and maintenance. organisation-wide implementation of these guidelines is critical to achieve changes in practice and patient health outcomes.ContextTo describe novel guideline implementation strategies used by a large US health care organisation to improve the care of obese and overweight adults.Description of Best PracticeAn evidence-based CPG was developed to address management strategies for a rapidly increasing number of obese and overweight patients. Interventions aimed at practitioner, patient and systems levels were tailored to facilitate implementation of CPG recommendations. Practitioner interventions included basic knowledge dissemination via electronic distribution of CPGs, presentation of CPGs at clinician champions’ meetings; and development of point-of-care job aids, such as in-clinic access to online CPGs and office prompts to refer obese patients to weight management classes. Patient-level interventions included proactive outreach for health education classes and telephone-based coaching; point-of-care educational publications; and after-visit summaries with weight management recommendations. Interventions at the systems level included proactive office encounter recording of patient BMI and exercise regimen; clinical performance goals; reporting of health outcomes of participants in weight management programmes; and, a physician continuing medical education (CME) course. Continued improvements in clinician/patient communication about weight, collection of patient weight information, and patient health outcomes have been observed.Lessons for Guideline Developers, Adaptors, Implementers, and/or UsersNovel approaches to integrating guidance on the management of obesity and overweight into practice can achieve significant changes in clinical practice and patient health outcomes.
072 Validation of the Guideline Implementability for Decision Excellence Model (Guide-M)
Authors: Brouwers M , Kastner M , Makarski J , Durocher L 
Published: August 2013

BackgroundWe developed a Guideline Implementability for Decision Excellence-Framework Model (GuIDE-M) based on the robust evidentiary base of a realist review on guideline attributes. GuIDE-M emerged as a conceptual representation of factors to facilitate the development of more implementable guidelines.ObjectivesValidity assessment of GuIDE-M with international guideline developers.MethodsWe assessed GuIDE-M using a stepwise validation process: Stage 1 involved consultation with a multi-disciplinary group of Canadian experts (including guideline research, psychology, management, and human factors engineering) to assess the sense and structure of the conceptual GuIDE-M. In Stage 2, 200 international guideline developers will assess GuIDE-M using an innovative online assessment platform, which includes an interactive video-based system to enable objective assessment of the model and its components.ResultsIn Stage 1, consultation with 10 multi-disciplinary experts informed major structural changes (e.g., addition of a 6th domain) and minor sense changes (e.g., collapsing like attributes) to the model. Stage 2 (in progress) will assess the organisation of the model according to Stage 1 findings: 1) To consider Stakeholder involvement, Evidence synthesis, Considered judgement and Feasibility in the development of guidelines; 2) and to communicate this content using effective Language and Format.DiscussionWe are applying an innovative stepwise process to rigorously validate and refine GuIDE-M internationally.Implications for Guideline Developers/UsersThis study represents a novel contribution to guideline developers and will offer a comprehensive, validated model that considers an exhaustive set of evidence-based factors to facilitate guideline uptake.
076 Enhancing the Acceptance and Implementation of GRADE Summary Tables for Evidence about Diagnostic Tests
Authors: Mustafa R , Wiercioch W , Brozek J , Lelgemann M , Buehler D , Garg A , Bossuyt P , Schünemann H 
Published: August 2013

BackgroundThe GRADE Working Group developed Summary tables adapted to summarise and present evidence from diagnostic test accuracy (DTA) systematic reviews.ObjectiveTo develop guidance on what information to include in these summary tables and to determine the best method(s) for presentation for different end users, including healthcare providers, systematic reviewers and guideline developers.MethodsWe presented a number of alternative summary tables to participants. We conducted questionnaires and one-on-one user testing interviews with target end users. We presented printed copies of summary tables and asked open-ended and 7-point Likert-scale questions to obtain information about users’ understanding and preferences.ResultsAll participants (n = 60) agreed that using summary tables to present results of DTA reviews is helpful. Presentation of several disease prevalence values was identified as a source of confusion. There was an overall preference for placement of sensitivity and specificity values inside summary tables to allow making a link to individual test results (TP, FN, TN, FP). A third of the participants read explanatory content in table footnotes. Two thirds of the participants noted that additional data, including adverse effects, costs, and treatment consequences, would be helpful for making appropriate conclusions and decisions about diagnostic tests.DiscussionAs results of DTA reviews are conceptually complicated, presenting the data in a clear, comprehensive, comprehensible way that is tailored to different end users is critical.Implications for Guideline Developers/UsersWe are developing a 3-layer approach, with varied content in summary tables of each layer tailored to the needs of different end users.
079 Formulating Guideline Recommendations about Important Clinical Questions with Low or Very Low Quality Evidence: The Case of Opioids for Chronic Pain
Authors: Harris J , Hegmann K , Weiss M 
Published: August 2013

BackgroundDeaths from prescription opioids now exceed those from street drugs or motor vehicle injuries in the US. Morbidity in has greatly increased as well. Rapid acceleration of opioid prescription for chronic non-cancer pain (CNCP) began in the mid-1990s with heavy marketing and support by opioid manufacturers. Some current recommendations advocate increased use of opioids despite a lack of quality evidence of long-term efficacy, considerable evidence of harms, and a tenuous understanding of CNCP.ObjectivesTo review the recommendation methodology used in cases of low quality evidence; to describe the process of recommendation development for opioid use for CNCPMethodsThe American College of Occupational and Environmental Medicine updated its systematic reviews and clinical practice guideline for the use of opioids for CNCP using critical appraisal and explicit panel methods. Panels consider population and clinical risk and benefit.ResultsCritical appraisal revealed low quality evidence. Most studies and many guideline panellists were funded by pharmaceutical companies. Harms were identified in observational studies.DiscussionAvailable guidelines tended to make vague recommendations that depended on clinician judgement. This panel therefore used methods to formulate recommendations that protect patients and the public, and a conservative and function-based approach to patient management.Implications for Guideline DevelopersGuidelines for areas in which evidence is low quality and the benefit to risk relationship is unclear should exercise caution in making recommendations, provide patient information, and recommend informed consent and careful patient management.
083 Developing and Implementing a Computerised Decision Support System for General Practice in the Netherlands
Authors: Bouma M , Lugtenberg M , Dorland R , Groot R de , Pasveer D , Kool T , Burgers J 
Published: August 2013

BackgroundThe Dutch College of General Practitioners (NHG) has produced almost 100 guidelines. Guideline adherence is on average 70–75%, but varies between general practitioners and between guideline recommendations. A computerised decision support system (CDSS) could improve guideline adherence.ObjectivesTo describe the development and implementation of a CDSS (NHGDoc) for general practice in the Netherlands.MethodsNHGDoc is a collaborative effort between the NHG and ExpertDoc, the company that developed the CDSS. Relevant recommendations of eight NHG guidelines were translated into algorithms, generating specific alerts. All alerts were reviewed and authorised by guideline developers of the NHG. NHGDoc was integrated in two out of seven electronic health record systems used in Dutch general practice. Since 2008, users were regularly interviewed regarding their experiences with NHGDoc.ResultsIntegration of eight guidelines in NHGDoc generated alerts in approximately 30% of all patient records. Alert frequency varied in accordance with disease prevalence. Currently, 1100 general practices (appr. 25% of Dutch general practices) are using NHGDoc. Users highly appreciated the content and lay out of the alerts, but preferred more control regarding the timing and frequency of the alerts.DiscussionNHGDoc is well received and appreciated in general practice. To enhance the success of NHGDoc the user-friendliness of the system could be further improved. Future research should provide evidence on the effectiveness of NHGDoc in terms of improving quality of care.Implications for Guideline DevelopersTo improve guideline adherence, CDSSs should have user-friendly designs, including options to personalise the decision support to meet the needs of individual users.