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Here, an in vitro characterization of a family of prazole derivatives that covalently bind to the C73 site on Tsg101 and assay their ability to inhibit viral particle production is presented. Structurally, increased steric bulk on the 4-pyridyl of the prazole expands the prazole site on the UEV domain toward the β-hairpin in the Ub-binding site and is coupled to increased inhibition of virus-like particle production in HIV-1. Increased bulk also increased toxicity, which is alleviated by increasing flexibility. Further, the formation of a novel secondary Tsg101 adduct for several of the tested compounds and the commercial drug lansoprazole. The secondary adduct involved the loss of the 4-pyridyl substituent to form an irreversible species, with implications for increasing the half-life of the active species or its specificity toward Tsg101 UEV. It is also determined that sulfide derivatives display effective viral inhibition, presumably through cellular sulfoxidation, allowing for delayed conversion within the cellular environment, and identify SARS-COV-2 as a target of prazole inhibition. These results open multiple avenues for the design of prazole derivatives for antiviral applications.

The Indonesian English Foreign Language (EFL) curriculum emphasizes using technology to enrich learning and teaching processes and encourages teachers to supplement the curriculum with context-relevant materials. However, little attention is given to the voices of teachers in implementing Computer Assisted Language Learning (CALL) for teaching English, especially how to elaborate technology, pedagogy, and content in Islamic schools where context and teacher agency remain underexplored. To fill this gap, we conducted professional development workshops to support teachers in Islamic school settings in their development of technological and pedagogical content knowledge. Twenty-one Islamic English teachers were involved in developing materials that supplemented the textbook used in Indonesian Islamic schools and at the same time introduced online and digital technologies into the EFL classroom. Drawing on Participatory Action Research (PAR), the data collected consisted of pre-and post-reflective journals, focus group discussion, field notes, participants assignment artefacts, and were analysed by thematic analysis. Our findings identified that the teachers enhanced their agency as material developers to utilize technology and pedagogy in EFL language teaching. Meanwhile, in terms of their content knowledge integration in TPACK in action, teachers integrated their cultural and religious background into materials, and this increased their confidence. This study also reports on challenges developing and sustaining TPACK.

This article reports on a project investigating the impact of a new framework for Readers Theatre on teachers' professional skills. Twenty-one Islamic secondary English school teachers engaged in collaborative professional development training to experience and give feedback on the framework. The article describes the participants' affective and critical responses and perspectives as well as opportunities and challenges related to the new framework in infusing peace values into EFL (English as a Foreign Language) materials. This study provides fresh insight into the potential of Readers Theatre for teaching peace and conflict prevention in the Indonesian Islamic EFL secondary school context.

Although previous work explained internal and external cultural challenges impacting academic integrity in the Muslim world, to date, no study has specifically examined the attempts and practices by the universities to address these challenges. The objective of this paper was to understand the actions taken by academics and institutions in the Muslim world to address, prevent breaches of academic integrity, and to recommend improvement of these practices. To capture institutional efforts and practices, relevant literature from 2010 to 2021 was reviewed to gather evidence of practices of academic integrity in higher education in the Muslim world. The findings suggest a framework that can be used for evaluation of current practices of academic integrity in the Muslim world, to go beyond plagiarism-focussed prevention, detection, and punishments.

Introduction: Enabling education programs, otherwise known as Foundation Studies or Preparatory programs, provide pathways for students typically under-represented in higher education. Students in Enabling programs often face distinct challenges in their induction to academic culture which can implicate them in cases of misconduct. This case study addresses a gap in the enabling literature reporting on how a culture of academic integrity can be developed for students and staff in these programs through an educative approach. Case description: This paper outlines how an educative approach to academic integrity is implemented within the Enabling programs of two Australian universities. Discussion and reflection: This case study reflects upon an approach which makes specific reference to the key elements of 'support', 'approach' and 'responsibility' as highlighted in Bretag and Mahmud's seminal paper. The paper reports a reduction in misconduct cases at the two institutions suggesting a positive correlation between the interventions and students' understanding of ethical academic practice. This study reflects upon practitioner experiences with academic integrity investigations to evaluate the effectiveness of this approach. Conclusions: The authors show that it is possible to ensure academic integrity practices and values are upheld within a supportive learning environment appropriate to a students' level of study.

Tsg101, a component of the endosomal sorting complex required for transport (ESCRT), is responsible for recognition of events requiring the machinery, as signaled by cargo tagging with ubiquitin (Ub), and for recruitment of downstream acting subunits to the site. Although much is known about the latter function, little is known about its role in the earlier event. The N-terminal domain of Tsg101 is a structural homologue of Ub conjugases (E2 enzymes) and the protein associates with Ub ligases (E3 enzymes) that regulate several cellular processes including virus budding. A pocket in the domain recognizes a motif, PT/SAP, that permits its recruitment. PT/SAP disruption makes budding dependent on Nedd4L E3 ligases. Using HIV-1 encoding a PT/SAP mutation that makes budding Nedd4L-dependent, we identified as critical for rescue the residues in the catalytic (HECT) domain of the E3 enzyme that lie in proximity to sites in Tsg101 that bind Ub non-covalently. Mutation of these residues impaired rescue by Nedd4L but the same mutations had no apparent effect in the context of a Nedd4 isomer, Nedd4-2s, whose N-terminal (C2) domain is naturally truncated, precluding C2-HECT auto-inhibition. Surprisingly, like small molecules that disrupt Tsg101 Ub-binding, small molecules that interfered with Nedd4 substrate recognition arrested budding at an early stage, supporting the conclusion that Tsg101–Ub–Nedd4 interaction promotes enzyme activation and regulates Nedd4 signaling for viral egress. Tsg101 regulation of E3 ligases may underlie its broad ability to function as an effector in various cellular activities, including viral particle assembly and budding. [ABSTRACT FROM AUTHOR]

BackgroundThe p6 region of the HIV-1 structural precursor polyprotein, Gag, contains two motifs, P7TAP11 and L35YPLXSL41, designated as late (L) domain-1 and -2, respectively. These motifs bind the ESCRT-I factor Tsg101 and the ESCRT adaptor Alix, respectively, and are critical for efficient budding of virus particles from the plasma membrane. L domain-2 is thought to be functionally redundant to PTAP. To identify possible other functions of L domain-2, we examined this motif in dominant viruses that emerged in a group of 14 women who had detectable levels of HIV-1 in both plasma and genital tract despite a history of current or previous antiretroviral therapy.ResultsRemarkably, variants possessing mutations or rare polymorphisms in the highly conserved L domain-2 were identified in seven of these women. A mutation in a conserved residue (S40A) that does not reduce Gag interaction with Alix and therefore did not reduce budding efficiency was further investigated. This mutation causes a simultaneous change in the Pol reading frame but exhibits little deficiency in Gag processing and virion maturation. Whether introduced into the HIV-1 NL4-3 strain genome or a model protease (PR) precursor, S40A reduced production of mature PR. This same mutation also led to high level detection of two extended forms of PR that were fairly stable compared to the WT in the presence of IDV at various concentrations; one of the extended forms was effective in trans processing even at micromolar IDV.ConclusionsOur results indicate that L domain-2, considered redundant in vitro, can undergo mutations in vivo that significantly alter PR function. These may contribute fitness benefits in both the absence and presence of PR inhibitor.

Abstract Background HIV-1 budding is directed primarily by two motifs in Gag p6 designated as late domain-1 and −2 that recruit ESCRT machinery by binding Tsg101 and Alix, respectively, and by poorly characterized determinants in the capsid (CA) domain. Here, we report that a conserved Gag p6 residue, S40, impacts budding mediated by all of these determinants. Results Whereas budding normally results in formation of single spherical particles ~100 nm in diameter and containing a characteristic electron-dense conical core, the substitution of Phe for S40, a change that does not alter the amino acids encoded in the overlapping pol reading frame, resulted in defective CA-SP1 cleavage, formation of strings of tethered particles or filopodia-like membrane protrusions containing Gag, and diminished infectious particle formation. The S40F-mediated release defects were exacerbated when the viral-encoded protease (PR) was inactivated or when L domain-1 function was disrupted or when budding was almost completely obliterated by the disruption of both L domain-1 and −2. S40F mutation also resulted in stronger Gag-Alix interaction, as detected by yeast 2-hybrid assay. Reducing Alix binding by mutational disruption of contact residues restored single particle release, implicating the perturbed Gag-Alix interaction in the aberrant budding events. Interestingly, introduction of S40F partially rescued the negative effects on budding of CA NTD mutations EE75,76AA and P99A, which both prevent membrane curvature and therefore block budding at an early stage. Conclusions The results indicate that the S40 residue is a novel determinant of HIV-1 egress that is most likely involved in regulation of a critical assembly event required for budding in the Tsg101-, Alix-, Nedd4- and CA N-terminal domain affected pathways.