Functional rescue of REP1 following treatment with PTC124 and novel derivative PTC-414 in human choroideremia fibroblasts and the nonsense-mediated zebrafish model

Choroideremia (CHM) is an X-linked chorioretinal dystrophy that is caused by mutations within a single gene, CHM Currently no effective treatment exists for these patients. Since over 30% of patients harbour nonsense mutations in CHM, nonsense suppression therapy using translational readthrough inducing drugs may provide functional rescue of REP1, thus attenuating progressive sight loss. Here, we employed two CHM model systems to systematically test the efficacy and safety of ataluren (PTC124) and its novel analog PTC-414: (1) the chm(ru848) zebrafish, the only nonsense mutation animal model of CHM harbouring a TAA nonsense mutation, and (2) a primary human fibroblast cell line from a CHM patient harbouring a TAG nonsense mutation. PTC124 or PTC-414 treatment of chm(ru848) embryos led to a ∼2.0-fold increase in survival, prevented the onset of retinal degeneration with reduced oxidative stress and apoptosis, increased rep1 protein by 23.1% (PTC124) and 17.2% (PTC-414) and restored biochemical function as confirmed through in vitro prenylation assays (98 ± 2% [PTC124] and 68 ± 5% [PTC-414]). In CHM(Y42X/y) fibroblasts, there was a recovery of prenylation activity following treatment with either PTC124 (42 ± 5%) or PTC-414 (36 ± 11%), although an increase in REP1 protein was not detected in these cells, in contrast to the zebrafish model. This comprehensive study on the use of PTC124 and PTC-414 as successful nonsense suppression agents for the treatment of CHM highlights the translational potential of these drugs for inherited retinal disease

Interaction of perceptual grouping and crossmodal temporal capture in tactile apparent-motion

Previous studies have shown that in tasks requiring participants to report the direction of apparent motion, task-irrelevant mono-beeps can "capture'' visual motion perception when the beeps occur temporally close to the visual stimuli. However, the contributions of the relative timing of multimodal events and the event structure, modulating uni- and/or crossmodal perceptual grouping, remain unclear. To examine this question and extend the investigation to the tactile modality, the current experiments presented tactile two-tap apparent-motion streams, with an SOA of 400 ms between successive, left-/right-hand middle-finger taps, accompanied by task-irrelevant, non-spatial auditory stimuli. The streams were shown for 90 seconds, and participants' task was to continuously report the perceived (left-or rightward) direction of tactile motion. In Experiment 1, each tactile stimulus was paired with an auditory beep, though odd-numbered taps were paired with an asynchronous beep, with audiotactile SOAs ranging from -75 ms to 75 ms. Perceived direction of tactile motion varied systematically with audiotactile SOA, indicative of a temporal-capture effect. In Experiment 2, two audiotactile SOAs-one short (75 ms), one long (325 ms)-were compared. The long-SOA condition preserved the crossmodal event structure (so the temporal-capture dynamics should have been similar to that in Experiment 1), but both beeps now occurred temporally close to the taps on one side (even-numbered taps). The two SOAs were found to produce opposite modulations of apparent motion, indicative of an influence of crossmodal grouping. In Experiment 3, only odd-numbered, but not even-numbered, taps were paired with auditory beeps. This abolished the temporal-capture effect and, instead, a dominant percept of apparent motion from the audiotactile side to the tactile-only side was observed independently of the SOA variation. These findings suggest that asymmetric crossmodal grouping leads to an attentional modulation of apparent motion, which inhibits crossmodal temporal-capture effects

Cultura digital e docência: possibilidades para a educação musical = Digital culture and teaching: possibilities for music education

Este artigo discute o modo como a Cultura Digital afeta a atuação docente na área da Educação Musical, problematizando o uso de recursos tecnológicos e seus desafios na prática pedagógica. A partir da reflexão sobre os conceitos de Musicalidade e Cultura Digital, o trabalho analisa um recorte sobre as transformações dos processos de aprendizagem e autoaprendizagem das práticas musicais subsidiadas pelas Novas Tecnologias Digitais (NTD) no contexto da Educação Musical. A metodologia foi constituída de revisão de literatura, questionário on-line e entrevista semiestruturada com músicos profissionais experientes, cujos dados coletados foram analisados na perspectiva quali-quantitativa. Os resultados demonstram que a Cultura Digital permeia as práticas pedagógicas do planejamento e pesquisa de materiais à implementação de recursos de aprendizagem e autoaprendizagem. A pesquisa aponta desafios da docência no campo da Educação Musical, considerando o descompasso constatado entre a fluência na Cultura Digital dos estudantes e as limitações materiais de infraestrutura, de conectividade e de formação inicial e continuada de professores. Defende que a Educação Musical se beneficia com o emprego das novas tecnologias digitais, porém exige uma mudança paradigmática ao entender o estudante como um ser integral e protagonista do processo educativo-musical, cuja bagagem tecnológica pode ser crucial no processo educativo

Cultura digital y docencia : posibilidades para la educación musical

Este artigo discute o modo como a Cultura Digital afeta a atuação docente na área da Educação Musical, problematizando o uso de recursos tecnológicos e seus desafios na prática pedagógica. A partir da reflexão sobre os conceitos de Musicalidade e Cultura Digital, o trabalho analisa um recorte sobre as transformações dos processos de aprendizagem e autoaprendizagem das práticas musicais subsidiadas pelas Novas Tecnologias Digitais (NTD) no contexto da Educação Musical. A metodologia foi constituída de revisão de literatura, questionário on-line e entrevista semiestruturada com músicos profissionais experientes, cujos dados coletados foram analisados na perspectiva quali-quantitativa. Os resultados demonstram que a Cultura Digital permeia as práticas pedagógicas do planejamento e pesquisa de materiais à implementação de recursos de aprendizagem e autoaprendizagem. A pesquisa aponta desafios da docência no campo da Educação Musical, considerando o descompasso constatado entre a fluência na Cultura Digital dos estudantes e as limitações materiais de infraestrutura, de conectividade e de formação inicial e continuada de professores. Defende que a Educação Musical se beneficia com o emprego das novas tecnologias digitais, porém exige uma mudança paradigmática ao entender o estudante como um ser integral e protagonista do processo educativo-musical, cuja bagagem tecnológica pode ser crucial no processo educativo.This article discusses how Digital Culture affects teachers’ work in the field of Music Education, debating the use of technological resources and their challenges in pedagogical practice. Reflecting upon the concepts of Musicality and Digital Culture, the work analyzes a focus on the change in processes of learning and self-learning of music practices assisted by New Digital Technologies (NDT) in the context of Musical Education. The methodology consisted of literature review, on-line questionnaire, and semi-structured interviews with experienced professional musicians, and data were analyzed under the qualitativequantitative perspective. The results demonstrate that the Digital Culture pervades pedagogical practices from planning and research of materials to implementation of learning and self-learning resources. The research points out challenges to teaching in the field of Music Education, considering the gap between students’ fluency in the Digital Culture and the material limitations of infrastructure, connectivity, and initial and continuing teacher training. It is argued that Music Education benefits from new digital technologies but requires a paradigmatic change to understand students as whole beings and subjects of the musical-educational process, whose technological experience may be crucial in the educational process.Este artículo discute el modo como la Cultura Digital afecta la actuación docente en el área de la Educación Musical, problematizando el uso de recursos tecnológicos y sus desafíos en la práctica pedagógica. A partir de la reflexión sobre los conceptos de Musicalidad y Cultura Digital, el artículo analiza un recorte sobre las transformaciones de los procesos de aprendizaje y autoaprendizaje de las prácticas musicales subsidiadas por las Nuevas Tecnologías Digitales (NTD) en el contexto de la Educación Musical. La metodología fue constituida de revisión de literatura, cuestionario en línea y entrevista semiestructurada con músicos profesionales experimentados, cuyos datos recolectados fueron analizados en la perspectiva cualitativa y cuantitativa. Los resultados demuestran que la Cultura Digitalpermea las prácticas pedagógicas de la planificación e investigación de materiales a la implementación de recursos de aprendizaje y autoaprendizaje. La investigación apunta desafíos de la docencia en el campo de la Educación Musical, considerando el descompaso constatado entre la fluidez en la Cultura Digital de los estudiantes y las limitaciones materiales de infraestructura, de conectividad y de formación inicial y continuada de profesores. Defiende que la Educación Musical se beneficia con el empleo de las nuevas tecnologías digitales, pero exige un cambio paradigmático al entender al estudiante como un ser integral y protagonista del proceso educativo-musical, cuyo equipaje tecnológico puede ser crucial en el proceso educativo

Human Musicality as seen from a Cognitive-Evolutionist Prism: From Homo sapiens to Homo digitalis

Este trabalho visa discutir a musicalidade humana sob um prisma cognitivo-evolucionista. Busca refletir acerca da complexidade da manifestação musical num panorama que articula os períodos transcorridos entre os tempos remotos da espécie Homo sapiens até a sua interação na era da Cultura Digital. As reflexões são fundamentadas em pesquisas que mostram um fazer musical ininterrupto, que inicia nos primórdios da humanidade e segue significativo até o presente momento, sofrendo, contudo, transformações que vêm sendo aceleradas pelas Novas Tecnologias Digitais (NTD). Esse cenário implica, também, mudanças na construção da performance musical, bem como nos conceitos de aprendizagem e autoaprendizagem de música.This paper discusses human musicality from a cognitive-evolutionist perspective. It reflects on the complexity of music manifestation in a panorama that articulates the periods between the remote times of the Homo sapiens species to their interaction in the era of Digital Culture. The reflections are based on research that shows uninterrupted music making that starts at the beginnings of humanity and continues significantly until the present moment, undergoing, however, transformations that are being accelerated by new digital technologies. This scenario also implies changes in the construction of musical performance, as well as in the concepts of learning and self-learning of music

No effect of auditory–visual spatial disparity on temporal recalibration

It is known that the brain adaptively recalibrates itself to small (∼100 ms) auditory–visual (AV) temporal asynchronies so as to maintain intersensory temporal coherence. Here we explored whether spatial disparity between a sound and light affects AV temporal recalibration. Participants were exposed to a train of asynchronous AV stimulus pairs (sound-first or light-first) with sounds and lights emanating from either the same or a different location. Following a short exposure phase, participants were tested on an AV temporal order judgement (TOJ) task. Temporal recalibration manifested itself as a shift of subjective simultaneity in the direction of the adapted audiovisual lag. The shift was equally big when exposure and test stimuli were presented from the same or different locations. These results provide strong evidence for the idea that spatial co-localisation is not a necessary constraint for intersensory pairing to occur

Fyn-Binding Protein (Fyb)/Slp-76–Associated Protein (Slap), Ena/Vasodilator-Stimulated Phosphoprotein (Vasp) Proteins and the Arp2/3 Complex Link T Cell Receptor (Tcr) Signaling to the Actin Cytoskeleton

T cell receptor (TCR)-driven activation of helper T cells induces a rapid polarization of their cytoskeleton towards bound antigen presenting cells (APCs). We have identified the Fyn- and SLP-76–associated protein Fyb/SLAP as a new ligand for Ena/ vasodilator-stimulated phosphoprotein (VASP) homology 1 (EVH1) domains. Upon TCR engagement, Fyb/SLAP localizes at the interface between T cells and anti-CD3–coated beads, where Evl, a member of the Ena/VASP family, Wiskott-Aldrich syndrome protein (WASP) and the Arp2/3 complex are also found. In addition, Fyb/SLAP is restricted to lamellipodia of spreading platelets. In activated T cells, Fyb/SLAP associates with Ena/VASP family proteins and is present within biochemical complexes containing WASP, Nck, and SLP-76. Inhibition of binding between Fyb/SLAP and Ena/VASP proteins or WASP and the Arp2/3 complex impairs TCR-dependent actin rearrangement, suggesting that these interactions play a key role in linking T cell signaling to remodeling of the actin cytoskeleton

[89Zr]Oxinate4 for long-term in vivo cell tracking by positron emission tomography

Purpose 111In (typically as [111In]oxinate3) is a gold standard radiolabel for cell tracking in humans by scintigraphy. A long half-life positron-emitting radiolabel to serve the same purpose using positron emission tomography (PET) has long been sought. We aimed to develop an 89Zr PET tracer for cell labelling and compare it with [111In]oxinate3 single photon emission computed tomography (SPECT). Methods [89Zr]Oxinate4 was synthesised and its uptake and efflux were measured in vitro in three cell lines and in human leukocytes. The in vivo biodistribution of eGFP-5T33 murine myeloma cells labelled using [89Zr]oxinate4 or [111In]oxinate3 was monitored for up to 14 days. 89Zr retention by living radiolabelled eGFP-positive cells in vivo was monitored by FACS sorting of liver, spleen and bone marrow cells followed by gamma counting. Results Zr labelling was effective in all cell types with yields comparable with 111In labelling. Retention of 89Zr in cells in vitro after 24 h was significantly better (range 71 to >90 %) than 111In (43–52 %). eGFP-5T33 cells in vivo showed the same early biodistribution whether labelled with 111In or 89Zr (initial pulmonary accumulation followed by migration to liver, spleen and bone marrow), but later translocation of radioactivity to kidneys was much greater for 111In. In liver, spleen and bone marrow at least 92 % of 89Zr remained associated with eGFP-positive cells after 7 days in vivo. Conclusion [89Zr]Oxinate4 offers a potential solution to the emerging need for a long half-life PET tracer for cell tracking in vivo and deserves further evaluation of its effects on survival and behaviour of different cell types

Lack of phenotypic and evolutionary cross-resistance against parasitoids and pathogens in Drosophila melanogaster

BackgroundWhen organisms are attacked by multiple natural enemies, the evolution of a resistance mechanism to one natural enemy will be influenced by the degree of cross-resistance to another natural enemy. Cross-resistance can be positive, when a resistance mechanism against one natural enemy also offers resistance to another; or negative, in the form of a trade-off, when an increase in resistance against one natural enemy results in a decrease in resistance against another. Using Drosophila melanogaster, an important model system for the evolution of invertebrate immunity, we test for the existence of cross-resistance against parasites and pathogens, at both a phenotypic and evolutionary level.MethodsWe used a field strain of D. melanogaster to test whether surviving parasitism by the parasitoid Asobara tabida has an effect on the resistance against Beauveria bassiana, an entomopathogenic fungus; and whether infection with the microsporidian Tubulinosema kingi has an effect on the resistance against A. tabida. We used lines selected for increased resistance to A. tabida to test whether increased parasitoid resistance has an effect on resistance against B. bassiana and T. kingi. We used lines selected for increased tolerance against B. bassiana to test whether increased fungal resistance has an effect on resistance against A. tabida.Results/ConclusionsWe found no positive cross-resistance or trade-offs in the resistance to parasites and pathogens. This is an important finding, given the use of D. melanogaster as a model system for the evolution of invertebrate immunity. The lack of any cross-resistance to parasites and pathogens, at both the phenotypic and the evolutionary level, suggests that evolution of resistance against one class of natural enemies is largely independent of evolution of resistance against the other

A nurse-led, preventive, psychological intervention to reduce PTSD symptom severity in critically ill patients: the POPPI feasibility study and cluster RCT

BACKGROUND: High numbers of patients experience severe acute stress in critical care units. Acute stress has been linked to post-critical care psychological morbidity, including post-traumatic stress disorder (PTSD). Previously, a preventive, complex psychological intervention [Psychological Outcomes following a nurse-led Preventative Psychological Intervention for critically ill patients (POPPI)] was developed by this research team, to be led by nurses, to reduce the development of PTSD symptom severity at 6 months. OBJECTIVES: The objectives were to (1) standardise and refine the POPPI intervention, and, if feasible, (2) evaluate it in a cluster randomised clinical trial (RCT). DESIGN: Two designs were used – (1) two feasibility studies to test the delivery and acceptability (to patients and staff) of the intervention, education package and support tools, and to test the trial procedures (i.e. recruitment and retention), and (2) a multicentre, parallel-group, cluster RCT with a baseline period and staggered roll-out of the intervention. SETTING: This study was set in NHS adult, general critical care units. PARTICIPANTS: The participants were adult patients who were > 48 hours in a critical care unit, receiving level 3 care and able to consent. INTERVENTIONS: The intervention comprised three elements – (1) creating a therapeutic environment in critical care, (2) three stress support sessions for patients identified as acutely stressed and (3) a relaxation and recovery programme for patients identified as acutely stressed. MAIN OUTCOMES MEASURES: Primary outcome – patient-reported symptom severity using the PTSD Symptom Scale – Self Report (PSS-SR) questionnaire (to measure clinical effectiveness) and incremental costs, quality-adjusted life-years (QALYs) and net monetary benefit at 6 months (to measure cost-effectiveness). Secondary outcomes – days alive and free from sedation to day 30; duration of critical care unit stay; PSS-SR score of > 18 points; depression, anxiety and health-related quality of life at 6 months; and lifetime cost-effectiveness. RESULTS: (1) A total of 127 participants were recruited to the intervention feasibility study from two sites and 86 were recruited to the RCT procedures feasibility study from another two sites. The education package, support tools and intervention were refined. (2) A total of 24 sites were randomised to the intervention or control arms. A total of 1458 participants were recruited. Twelve sites delivered the intervention during the intervention period: > 80% of patients received two or more stress support sessions and all 12 sites achieved the target of > 80% of clinical staff completing the POPPI online training. There was, however, variation in delivery across sites. There was little difference between baseline and intervention periods in the development of PTSD symptom severity (measured by mean PSS-SR score) at 6 months for surviving patients in either the intervention or the control group: treatment effect estimate −0.03, 95% confidence interval (CI) −2.58 to 2.52; p = 0.98. On average, the intervention decreased costs and slightly improved QALYs, leading to a positive incremental net benefit at 6 months (£835, 95% CI −£4322 to £5992), but with considerable statistical uncertainty surrounding these results. There were no significant differences between the groups in any of the secondary outcomes or in the prespecified subgroup analyses. LIMITATIONS: There was a risk of bias because different consent processes were used and as a result of the lack of blinding, which was mitigated as far as possible within the study design. The intervention started later than anticipated. Patients were not routinely monitored for delirium. CONCLUSIONS: Among level 3 patients who stayed > 48 hours in critical care, the delivery of a preventive, complex psychological intervention, led by nurses, did not reduce the development of PTSD symptom severity at 6 months, when compared with usual care. FUTURE WORK: Prior to development and evaluation of subsequent psychological interventions, there is much to learn from post hoc analyses of the cluster RCT rich quantitative and qualitative data. TRIAL REGISTRATION: This trial is registered as ISRCTN61088114 and ISRCTN53448131. FUNDING: This project was funded by the National Institute for Health Research (NIHR) Health Services and Delivery Research programme and will be published in full in Health Services and Delivery Research; Vol. 23, No. 30. See the NIHR Journals Library website for further project information