67 research outputs found

    Positive Selection and Increased Antiviral Activity Associated with the PARP-Containing Isoform of Human Zinc-Finger Antiviral Protein

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    Intrinsic immunity relies on specific recognition of viral epitopes to mount a cell-autonomous defense against viral infections. Viral recognition determinants in intrinsic immunity genes are expected to evolve rapidly as host genes adapt to changing viruses, resulting in a signature of adaptive evolution. Zinc-finger antiviral protein (ZAP) from rats was discovered to be an intrinsic immunity gene that can restrict murine leukemia virus, and certain alphaviruses and filoviruses. Here, we used an approach combining molecular evolution and cellular infectivity assays to address whether ZAP also acts as a restriction factor in primates, and to pinpoint which protein domains may directly interact with the virus. We find that ZAP has evolved under positive selection throughout primate evolution. Recurrent positive selection is only found in the poly(ADP-ribose) polymerase (PARP)–like domain present in a longer human ZAP isoform. This PARP-like domain was not present in the previously identified and tested rat ZAP gene. Using infectivity assays, we found that the longer isoform of ZAP that contains the PARP-like domain is a stronger suppressor of murine leukemia virus expression and Semliki forest virus infection. Our study thus finds that human ZAP encodes a potent antiviral activity against alphaviruses. The striking congruence between our evolutionary predictions and cellular infectivity assays strongly validates such a combined approach to study intrinsic immunity genes

    Diversifying selection and functional analysis of interleukin-4 suggests antagonism-driven evolution at receptor-binding interfaces

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    <p>Abstract</p> <p>Background</p> <p>Interleukin-4 (IL4) is a secreted immunoregulatory cytokine critically involved in host protection from parasitic helminths <abbrgrp><abbr bid="B1">1</abbr></abbrgrp>. Reasoning that helminths may have evolved mechanisms to antagonize IL4 to maximize their dispersal, we explored mammalian IL4 evolution.</p> <p>Results</p> <p>This analysis revealed evidence of diversifying selection at 15 residues, clustered in epitopes responsible for IL4 binding to its Type I and Type II receptors. Such a striking signature of selective pressure suggested either recurrent episodes of pathogen antagonism or ligand/receptor co-evolution. To test the latter possibility, we performed detailed functional analysis of IL4 allotypes expressed by <it>Mus musculus musculus </it>and <it>Mus musculus castaneus</it>, which happen to differ at 5 residues (including three at positively selected sites) in and adjacent to the site 1 epitope that binds the IL4Rα subunit shared by the Type I and Type II IL4 receptors. We show that this intra-species variation affects the ability of IL4 neither to bind IL4 receptor alpha (IL4Rα) nor to signal biological responses through its Type I receptor.</p> <p>Conclusions</p> <p>Our results -- reminiscent of clustered positively selected sites revealing functionally important residues at host-virus interaction interfaces -- are consistent with IL4 having evolved to avoid recurrent pathogen antagonism, while maintaining the capacity to bind and signal through its cognate receptor. This work exposes what may be a general feature of evolutionary conflicts fought by pathogen antagonists at host protein-protein interaction interfaces involved in immune signaling: the emergence of receptor-binding ligand epitopes capable of buffering amino acid variation.</p

    Supervising EBT: What Content Do Workplace-Based Supervisors Cover and What Techniques Do They Use?

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    Workplace-based clinical supervision in public mental health is an underutilized resource for supporting evidence- based treatments (EBTs) [1], despite the fact that supervisors may offer a cost-effective way to support clinician fidelity to EBT. Very little, however, is known about the content and techniques used by workplace-based supervisors [2]; particularly in the context of EBT implementation [3]

    Objective Coding of Content and Techniques in Workplace-Based Supervision of an EBT in Public Mental Health

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    BACKGROUND: Workplace-based clinical supervision as an implementation strategy to support evidence-based treatment (EBT) in public mental health has received limited research attention. A commonly provided infrastructure support, it may offer a relatively cost-neutral implementation strategy for organizations. However, research has not objectively examined workplace-based supervision of EBT and specifically how it might differ from EBT supervision provided in efficacy and effectiveness trials. METHODS: Data come from a descriptive study of supervision in the context of a state-funded EBT implementation effort. Verbal interactions from audio recordings of 438 supervision sessions between 28 supervisors and 70 clinicians from 17 public mental health organizations (in 23 offices) were objectively coded for presence and intensity coverage of 29 supervision strategies (16 content and 13 technique items), duration, and temporal focus. Random effects mixed models estimated proportion of variance in content and techniques attributable to the supervisor and clinician levels. RESULTS: Interrater reliability among coders was excellent. EBT cases averaged 12.4 min of supervision per session. Intensity of coverage for EBT content varied, with some discussed frequently at medium or high intensity (exposure) and others infrequently discussed or discussed only at low intensity (behavior management; assigning/reviewing client homework). Other than fidelity assessment, supervision techniques common in treatment trials (e.g., reviewing actual practice, behavioral rehearsal) were used rarely or primarily at low intensity. In general, EBT content clustered more at the clinician level; different techniques clustered at either the clinician or supervisor level. CONCLUSIONS: Workplace-based clinical supervision may be a feasible implementation strategy for supporting EBT implementation, yet it differs from supervision in treatment trials. Time allotted per case is limited, compressing time for EBT coverage. Techniques that involve observation of clinician skills are rarely used. Workplace-based supervision content appears to be tailored to individual clinicians and driven to some degree by the individual supervisor. Our findings point to areas for intervention to enhance the potential of workplace-based supervision for implementation effectiveness. TRIAL REGISTRATION: NCT01800266 , Clinical Trials, Retrospectively Registered (for this descriptive study; registration prior to any intervention [part of phase II RCT, this manuscript is only phase I descriptive results])

    Use of Sperm In Vitro Capacitation and Flow Cytometry to Estimate Bull Fertility

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    Study Description: Frozen-thawed semen from five bulls previously identified as high (48.1% and 47.7%, bulls A and B, respectively), intermediary (45.5%, bull C) or low (43.1% and 40.7%, bulls D and E, respectively) fertility, based on pregnancy per AI, were evaluated with several laboratory measures. Measures included total motility, sperm plasma membrane integrity (viability), acrosome integrity, reactive oxygen species (ROS), mitochondrial membrane energy potential (mito-potential), zinc signatures (signatures 1 to 4), and CD9 protein populations at pre-wash, post-wash, h 0 (diluted with non-capacitation media), and at 0, 3, 6, and 24 h after dilution with capacitation media and incubation at 37 ºC. Data were analyzed using the GLIMMIX procedure of SAS for repeated measures with bull, time, and the interaction as fixed effects. Bull by time interaction was significant (P ≤ 0.01) for total motility and viability. There tended (P = 0.06) to be a bull by time interaction for zinc signatures 1 + 2 combined. There was a significant effect of bull (P ≤ 0.03) for viability, viable sperm with disrupted acrosome, zinc signatures 1, 2, and 1 + 2, viable CD9- (CD9 negative), and dead CD9+ (CD9 positive). High and intermediary field fertility bulls had greater (P ≤ 0.04) percentages of viable sperm, zinc signature 2, and zinc signature 1 + 2 compared to low fertility bulls. High and intermediary fertility bulls had decreased (P ≤ 0.05) percentage of dead CD9+ compared to low fertility bulls. There was or tended to be a positive correlation between pregnancy per AI and viability (P = 0.10; r = 0.81), zinc signature 2 (P = 0.04; r = 0.89), and zinc signature 1 + 2 (P = 0.10; r = 0.80)
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