Diagnosis of Alzheimer Disease and Tauopathies on Whole-Slide Histopathology Images Using a Weakly Supervised Deep Learning Algorithm.

Neuropathologic assessment during autopsy is the gold standard for diagnosing neurodegenerative disorders. Neurodegenerative conditions, such as Alzheimer disease (AD) neuropathological change, are a continuous process from normal aging rather than categorical; therefore, diagnosing neurodegenerative disorders is a complicated task. We aimed to develop a pipeline for diagnosing AD and other tauopathies, including corticobasal degeneration (CBD), globular glial tauopathy, Pick disease, and progressive supranuclear palsy. We used a weakly supervised deep learning-based approach called clustering-constrained-attention multiple-instance learning (CLAM) on the whole-slide images (WSIs) of patients with AD (n = 30), CBD (n = 20), globular glial tauopathy (n = 10), Pick disease (n = 20), and progressive supranuclear palsy (n = 20), as well as nontauopathy controls (n = 21). Three sections (A: motor cortex; B: cingulate gyrus and superior frontal gyrus; and C: corpus striatum) that had been immunostained for phosphorylated tau were scanned and converted to WSIs. We evaluated 3 models (classic multiple-instance learning, single-attention-branch CLAM, and multiattention-branch CLAM) using 5-fold cross-validation. Attention-based interpretation analysis was performed to identify the morphologic features contributing to the classification. Within highly attended regions, we also augmented gradient-weighted class activation mapping to the model to visualize cellular-level evidence of the model's decisions. The multiattention-branch CLAM model using section B achieved the highest area under the curve (0.970 ± 0.037) and diagnostic accuracy (0.873 ± 0.087). A heatmap showed the highest attention in the gray matter of the superior frontal gyrus in patients with AD and the white matter of the cingulate gyrus in patients with CBD. Gradient-weighted class activation mapping showed the highest attention in characteristic tau lesions for each disease (eg, numerous tau-positive threads in the white matter inclusions for CBD). Our findings support the feasibility of deep learning-based approaches for the classification of neurodegenerative disorders on WSIs. Further investigation of this method, focusing on clinicopathologic correlations, is warranted.

An analysis of a video education campaign on COVID-19 for healthcare workers in Africa delivered through Facebook.

INTRODUCTION: The COVID-19 pandemic required rapid dissemination of accurate information across the world to both healthcare workers and the general public. Social media represents an opportunity to undertake this. The aim of this study was to analyse a healthcare worker education campaign in Africa delivered through the social media platform Facebook and discuss the feasibility of this approach for future healthcare workers and public health campaigns. METHODS: The campaign ran from June 2020 to January 2021. The Facebook Ad Manager suite was used to extract data in July 2021. Videos were analysed for total and individual video reach, impressions, 3-second video plays, 50% plays and 100% plays. The geographic use of the videos and age and gender breakdown was also analysed. RESULTS: Total reach of the Facebook campaign was 6,356,846 and total impressions was 12,767,118. The video with the highest reach was 'Hand washing steps for health workers' with a reach of 1,479,603. The total campaign 3-second plays were 2,189,460 decreasing to 77,120 for 100% play duration. DISCUSSION: Facebook advertising campaigns may have the ability to reach large populations and achieve a range of engagement outcomes that would be more cost effective and have greater reach when compared with traditional media. The outcome of this campaign has shown the potential of social media's use in public health information, medical education and professional development.

The availability of general practice in Ireland's Mid-West Region: does the 'Inverse Care Law' still apply?

INTRODUCTION: The 'Inverse Care Law' suggests the availability of good medical care tends to vary inversely with the needs of the local population. Dr Julian Tudor Hart's observations related to lack of access to care for those in both socially deprived and geographically remote areas. In this study, we aim to examine if the 'Inverse Care Law' is still relevant to GP service provision in the Mid-West of Ireland. METHODS: GP clinic locations in Limerick and Clare were identified using the Health Service Executive (HSE) Service Finder and geocoded. GeoHive.ie was used to determine Electoral District (ED) centroids across the Mid-West. The shortest linear distance to a GP clinic was calculated for each ED. PobalMaps.ie was used to determine population and social deprivation scores of each ED. RESULTS: In total, 122 GP practices were identified across 324 EDs. The average travel distance to a GP clinic in the Mid-West is 4.7 km. Limerick City EDs had the smallest patient population per GP clinic and were all found to be within 1.5 km of a GP clinic. Proximity to GP clinics did not correlate with deprivation. However, by removing GP clinics from the analyses, it was possible to determine how vulnerable different areas (rural vs urban, deprived vs affluent) are to potential changes in GP clinic availability in the future. DISCUSSION: People living in urban areas such a Limerick City have improved geographic accessibility to GP clinics compared with their rural counterparts. However, within urban areas assessed, GP clinics were rarely found in deprived areas. Therefore, remote and urban-deprived areas are far more vulnerable to negative proximity effects secondary to practice closures, suggesting the principles of the 'Inverse Care Law' may still be active in the Mid-West of Ireland.

GPs at the Edge: a quantitative description of Irish Rural General Practice.

INTRODUCTION: Currently, more than 1.6 million Irish people live rurally. Rural populations in Ireland are older and have more health needs compared with younger urban areas. Meanwhile, since 1982, the proportion of general practices in rural areas has decreased by 10%. In this study, we look at new survey data to investigate the needs and challenges of rural general practice in Ireland. METHODS: This study will make use of survey responses from the 2021 membership survey by the Irish College of General Practitioners (ICGP). The anonymous, online, survey was sent by email to the ICGP membership in late 2021, with a series of questions pertaining to practice location, and prior experience of living and working in a rural area designed specifically for this project. A series of statistical tests will be undertaken as appropriate for the data. RESULTS: This study is ongoing; we aim to present data on the demographics of those working in rural general practice and related factors. DISCUSSION: Previous research has shown that people who grew up or trained in rural areas are more likely to work there after qualifying. As the analysis of this survey continues, it will be important to see if this pattern is evident here as well.

GPs at the Edge: GATE, a qualitative investigation into the experience of delivering care in Irish Rural General Practice.

INTRODUCTION: Over a third of Irish people live rurally. However, only a fifth of Irish general practices are located in rural communities and longstanding issues such as distance to other health services, professional isolation, and recruitment and retention of rural healthcare professionals (HCPs) threaten rural general practice's sustainability. This ongoing study seeks to understand what it is like to provide care to Ireland's rural and remote populations. METHODS: This is a qualitative study, consisting of semi-structured interviews with GPs and practice nurses working in practices that serve rural populations across Ireland. Topic guides were developed after a literature review and a series of pilot interviews. Interviews are scheduled to be finished in February 2022. RESULTS: This study is ongoing so results are yet to be finalised. Initial key themes include a great level of professional satisfaction that GPs and practice nurses experience from caring for entire families from the 'cradle to grave' and from the complex issues they face in practice. A rural general practice acts as the medical port-of-call for patients, with both practice nurses and GPs having experiences with emergency and pre-hospital medicine. A key difficulty identified is access to secondary and tertiary care services, with distance to services and high demand as the main barriers. DISCUSSION: Working in rural general practice gives HCPs great professional satisfaction but access to other health services remains a challenge. Final conclusions may be compared with other delegates' experiences.

Galactosylceramidase deficiency and pathological abnormalities in cerebral white matter of Krabbe disease.

Krabbe Disease (KD) is an autosomal recessive disorder that results from loss-of-function mutations in the GALC gene, which encodes lysosomal enzyme galactosylceramidase (GALC). Functional deficiency of GALC is toxic to myelin-producing cells, which leads to progressive demyelination in both the central and peripheral nervous systems. It is hypothesized that accumulation of psychosine, which can only be degraded by GALC, is a primary initiator of pathologic cascades. Despite the central role of GALC in KD pathomechanism, investigations of GALC deficiency at a protein level are largely absent, due in part, to the lack of sensitive antibodies in the field. Leveraging two custom antibodies that can detect GALC at endogenous levels, we demonstrated that GALC protein is predominantly localized to oligodendrocytes in cerebral white matter of an infant brain, consistent with its functional role in myelination. Mature GALC could also be quantitatively detected as a 26 kDa band by western blotting and correlated to enzyme activity in brain tissues. The p.Ile562Thr polymorphic variant, which is over-represented in the KD population, was associated with reduced mature GALC protein and activity. In three infantile KD cases, homozygous null mutations in GALC lead to deficiency in total GALC protein and activity. Interestingly, although GALC activity was absent, normal levels of total GALC protein were detected by a sandwich ELISA using our custom antibodies in a later-onset KD brain, which suggests that the assay has the potential to differentiate infantile- and later-onset KD cases. Among the infantile KD cases, we quantified a 5-fold increase in psychosine levels, and observed increased levels of acid ceramidase, a key enzyme for psychosine production, and hyperglycosylated lysosomal-associated membrane protein 1, a marker for lysosomal activation, in periventricular white matter, a major pathological brain region, when compared with age-matched normal controls. While near complete demyelination was observed in these cases, we quantified that an early-infantile case (age of death at 10 months) had about 3-fold increases in both globoid cells, a pathological hallmark for KD, and CD8-positive T lymphocytes, a pathological marker for multiple sclerosis, in the white matter when compared with a slower progressing infantile case (age of death at 21 months), which suggests a positive correlation between clinical severity and neuropathology. Taken together, our findings have advanced the understanding of GALC protein biology in the context of normal and KD brain white matter. We also revealed new neuropathological changes that may provide insights to understand KD pathogenesis.

Loss of Tmem106b exacerbates FTLD pathologies and causes motor deficits in progranulin-deficient mice.

Progranulin (PGRN) and transmembrane protein 106B (TMEM106B) are important lysosomal proteins implicated in frontotemporal lobar degeneration (FTLD) and other neurodegenerative disorders. Loss-of-function mutations in progranulin (GRN) are a common cause of FTLD, while TMEM106B variants have been shown to act as disease modifiers in FTLD. Overexpression of TMEM106B leads to lysosomal dysfunction, while loss of Tmem106b ameliorates lysosomal and FTLD-related pathologies in young Grn-/- mice, suggesting that lowering TMEM106B might be an attractive strategy for therapeutic treatment of FTLD-GRN. Here, we generate and characterize older Tmem106b-/- Grn-/- double knockout mice, which unexpectedly show severe motor deficits and spinal cord motor neuron and myelin loss, leading to paralysis and premature death at 11-12 months. Compared to Grn-/- , Tmem106b-/- Grn-/- mice have exacerbated FTLD-related pathologies, including microgliosis, astrogliosis, ubiquitin, and phospho-Tdp43 inclusions, as well as worsening of lysosomal and autophagic deficits. Our findings confirm a functional interaction between Tmem106b and Pgrn and underscore the need to rethink whether modulating TMEM106B levels is a viable therapeutic strategy.

Lessons learned from a pandemic: implications for a combined exercise and educational programme for medical students.

BACKGROUND: The 'MED-WELL' programme is a combined exercise and educational intervention designed to promote well-being among medical students and educate students about prescribing exercise as medicine in clinical practice. Due to COVID-19 public health restrictions of social distancing the 'MED-WELL' programme was offered online instead of in-person in 2021. The aim of this study is to compare the experiences of participants in the 'MED-WELL' programme online to those that previously participated in the same programme in-person to understand the student experience and optimize programme delivery. METHODS: Purposive sampling was used to recruit 20 participants to a qualitative study using semi-structured interviews. Ten study participants took part in the 'MED-WELL' programme when it was offered in-person, and the other ten study participants took part in the programme when it was offered online. All interviews were audio-recorded and transcribed using Microsoft Teams. A combined inductive and deductive approach was used for analysis. An inductive thematic analysis was utilized to categorize data into higher order codes, themes, and overarching themes. The theory of online learning provided the theoretical framework for a deductive approach. RESULTS: Analysis of the data produced five overarching themes: 'student-student', 'student-teacher', 'student-content', 'student-environment', and 'effects of a pandemic'. The first four themes detail distinct types of interaction that participants had with various entities of the 'MED-WELL' programme and the effects that these interactions had on participant experiences. 'Effects of a pandemic' refers to the context of delivering the 'MED-WELL' programme online during a pandemic and how this mode of delivery influenced participants and the programme. CONCLUSIONS: Optimizing the 'MED-WELL' programme relies on an understanding of how participants interact with different entities of the programme and are motivated to attend and engage. Participants tended to favour an in-person mode of delivery, however certain advantages of delivering the programme online were also identified. The findings from this study can be used to inform similar experiential and educational exercise interventions, and may help plan for potential future restrictions on in-person educational and exercise-based programmes.

Phosphorylated tau sites that are elevated in Alzheimer's disease fluid biomarkers are visualized in early neurofibrillary tangle maturity levels in the post mortem brain.

INTRODUCTION: Alzheimer's disease (AD) biomarkers are increasingly more reliable in predicting neuropathology. To facilitate interpretation of phosphorylated tau sites as an early fluid biomarker, we sought to characterize which neurofibrillary tangle maturity levels (pretangle, intermediary 1, mature tangle, intermediary 2, and ghost tangle) are recognized. METHODS: We queried the Florida Autopsied Multi-Ethnic (FLAME) cohort for cases ranging from Braak stages I through VI, excluding non-AD neuropathologies and tauopathies. Thioflavin-S staining was compared to immunohistochemical measures of phosphorylated threonine (pT) 181, pT205, pT217, and pT231 in two hippocampal subsectors across n = 24 cases. RESULTS: Each phosphorylated tau site immunohistochemically labeled early neurofibrillary tangle maturity levels compared to advanced levels recognized by thioflavin-S. Hippocampal burden generally increased with each Braak stage. DISCUSSION: These results provide neurobiologic evidence that these phosphorylated tau fluid biomarker sites are present during early neurofibrillary tangle maturity levels and may explain why these fluid biomarker measures are observed before symptom onset. HIGHLIGHTS: Immunohistochemical evaluation of four phosphorylated tau fluid biomarker sites. Earlier neurofibrillary tangle maturity levels recognized by phosphorylated tau in proline-rich region. Advanced tangle pathology is elevated in the subiculum compared to the cornu ammonis 1 of the hippocampus. Novel semi-quantitative frequency to calculate tangle maturity frequency.

Loss of TMEM106B leads to myelination deficits: implications for frontotemporal dementia treatment strategies.

Genetic variants that define two distinct haplotypes at the TMEM106B locus have been implicated in multiple neurodegenerative diseases and in healthy brain ageing. In frontotemporal dementia (FTD), the high expressing TMEM106B risk haplotype was shown to increase susceptibility for FTD with TDP-43 inclusions (FTD-TDP) and to modify disease penetrance in progranulin mutation carriers (FTD-GRN). To elucidate the biological function of TMEM106B and determine whether lowering TMEM106B may be a viable therapeutic strategy, we performed brain transcriptomic analyses in 8-month-old animals from our recently developed Tmem106b-/- mouse model. We included 10 Tmem106b+/+ (wild-type), 10 Tmem106b+/- and 10 Tmem106-/- mice. The most differentially expressed genes (153 downregulated and 60 upregulated) were identified between Tmem106b-/- and wild-type animals, with an enrichment for genes implicated in myelination-related cellular processes including axon ensheathment and oligodendrocyte differentiation. Co-expression analysis also revealed that the most downregulated group of correlated genes was enriched for myelination-related processes. We further detected a significant loss of OLIG2-positive cells in the corpus callosum of Tmem106b-/- mice, which was present already in young animals (21 days) and persisted until old age (23 months), without worsening. Quantitative polymerase chain reaction revealed a reduction of differentiated but not undifferentiated oligodendrocytes cellular markers. While no obvious changes in myelin were observed at the ultrastructure levels in unchallenged animals, treatment with cuprizone revealed that Tmem106b-/- mice are more susceptible to cuprizone-induced demyelination and have a reduced capacity to remyelinate, a finding which we were able to replicate in a newly generated Tmem106b CRISPR/cas9 knock-out mouse model. Finally, using a TMEM106B HeLa knock-out cell line and primary cultured oligodendrocytes, we determined that loss of TMEM106B leads to abnormalities in the distribution of lysosomes and PLP1. Together these findings reveal an important function for TMEM106B in myelination with possible consequences for therapeutic strategies aimed at lowering TMEM106B levels.

Communication between primary and secondary care: deficits and danger.

BACKGROUND: Timely and accurate communication between primary and secondary care is essential for delivering high-quality patient care. OBJECTIVE: The aim of this study is to evaluate the content contained in both referral and response letters between primary and secondary care and measure this against the recommended national guidelines. METHODS: Using an observational design, senior medical students and their general practice supervisors applied practice management software to identify 100 randomly selected adults, aged greater than 50 years, from a generated list of consults over a 2-year period (2013-2015). All data included in referral and response letters for these adults were examined and compared with the gold standard templates that were informed by international guidelines. RESULTS: Data from 3293 referral letters and 2468 response letters from 68 general practices and 17 hospitals were analysed. The median time that had elapsed between a patient being referred and receiving a response letter was 4 weeks, ranging from 1 week for Emergency Department referral letters to 7 weeks for orthopaedic surgery referral letters. Referral letters included the reason for referral (98%), history of complaint (90%) and current medications (82%). Less commonly included were management prior to referral (65%) and medication allergies (57%). The majority of response letters included information on investigations (73%), results (70%) and follow-up plan (85%). Less commonly, response letters included medication changes (30%), medication lists (33%) and secondary diagnoses (13%). CONCLUSIONS: Future research should be aimed at developing robust strategies to addressing communication gaps reported in this study.

The University of Limerick Education and Research Network for General Practice (ULEARN-GP): practice characteristics and general practitioner perspectives.

BACKGROUND: A well-functioning general practice sector that has a strong research component is recognised as a key foundation of any modern health system. General practitioners (GPs) are more likely to collaborate in research if they are part of an established research network. The primary aims of this study are to describe Ireland's newest general practice-based research network and to analyse the perspectives of the network's members on research engagement. METHOD: A survey was sent to all GPs participating in the network in order to document practice characteristics so that this research network's profile could be compared to other national profiles of Irish general practice. In depth interviews were then conducted and analysed thematically to explore the experiences and views of a selection of these GPs on research engagement. RESULTS: All 134 GPs responded to the survey. Practices have similar characteristics to the national profile in terms of location, size, computerisation, type of premises and out of hours arrangements. Twenty-two GPs were interviewed and the resulting data was categorised into subthemes and four related overarching themes: GPs described catalysts for research in their practices, the need for coherence in how research is understood in this context, systems failures, whereby the current health system design is prohibitive of GP participation and aspirations for a better future. CONCLUSION: This study has demonstrated that the research network under examination is representative of current trends in Irish general practice. It has elucidated a better understanding of factors that need to be addressed in order to encourage more GPs to engage in the research process.

A feasibility study of an exercise intervention to educate and promote health and well-being among medical students: the 'MED-WELL' programme.

BACKGROUND: Medical School programme workloads challenge the physical and mental health of students particularly in compressed graduate entry programmes. There is evidence that physical activity (PA) can improve holistic care and help maintain wellness among medical students. We tested the feasibility of introducing an exercise programme to the medical school curriculum which would educate and promote health and well-being among its students. METHODS: This study was conducted in a single graduate entry medical school at the University of Limerick (UL). The 'MED-WELL' programme was a six-week programme of 1 hour-long weekly sessions, each involving a different type of PA (45 min). These sessions were prefaced by an interactive lecture about how to incorporate exercise theory into daily medical practice (15 min). The study was conducted in a single graduate entry medical school at UL and involved year one and year two graduate entry medical students. Three parameters were used to test feasibility: 1. Recruitment and retention of participants, 2. Acceptability of the programme and 3. Efficacy in terms of health and well-being. The latter was assessed by administering questionnaires pre and post the intervention. The questionnaires used the following validated measurement scales: EQ-VAS; WHO-5 Well-Being Index; 3-item Loneliness Scale; Social Support Measure 3-item scale. Free text boxes also encouraged participants to discuss the merits of the programme. RESULTS: In total, 26% (74/286 students) participated in the programme. Of those who participated, 69 students (93%) attended one or more sessions of the programme and completed questionnaires at baseline and at follow-up. Significant improvements were seen in scores after the programme in the WHO-5 Well-Being Index which increased from 63.2 (95%CI: 48-78.4) to 67.5 (95%CI: 55.1-79.9); (P < 0.01), the sleep scale which increased from 3.1 (95%CI: 2.2-4.0) to 3.5 (95%CI: 2.5-4.5); (P < 0.001), and the loneliness scale which decreased from 4.1 (95%CI: 2.7-5.5) to 3.5 (95%CI: 2.5-4.5); (P < 0.005). Students level of PA during a typical week also increased from 3.7 (95%CI: 2.1-5.4) to 4.0 (95%CI, 3.5-4.5); (P < 0.05). CONCLUSION: This study has shown it is feasible to deliver this programme in a medical school's curriculum. The programme seems to be of benefit and is acceptable to students. Well-designed randomised controlled trials are needed to measure outcomes, durability of effect, and cost effectiveness.

A qualitative comparison of high and low adherers with apparent treatment-resistant hypertension.

Poor adherence is a leading cause of apparent resistance to antihypertensive treatment. Recent empirical research has investigated predictors of adherence for primary care patients who are apparently resistant to treatment; however, questions remain regarding the variability in adherence behaviour among this group. This study aimed to investigate factors that may elucidate medication adherence among patients with apparent treatment-resistant hypertension (aTRH) using qualitative methods. Fourteen semi-structured interviews were conducted with patients undergoing treatment for aTRH in primary care in the West of Ireland. Patients who self-reported both high and low adherence in a previous quantitative study were purposively sampled. Data were analysed using thematic analysis. A public and patient involvement research group were active partners in developing the study protocol and interview topic guide. Three major themes were identified: beliefs about treatment, habits and routine, and health and health systems. High adherers reported favourable beliefs about antihypertensive treatment that had been validated by experience with taking the treatment over time, described strong medication-taking habits and stable routines, and positive relations with their GP. Low adherers expressed less coherence in their beliefs and used less effective strategies to support their medication-taking in daily life. The current findings are consistent with qualitative studies of adherence in other chronic conditions. Results reflect the difficulty for healthcare practitioners in identifying adherent versus non-adherent patients via conversation, and highlight the importance of accurate adherence assessment. Inception studies may provide an opportunity to better understand adherence behaviour across the illness trajectory.

An acetylation-phosphorylation switch that regulates tau aggregation propensity and function.

The aberrant accumulation of tau protein is a pathological hallmark of a class of neurodegenerative diseases known as tauopathies, including Alzheimer's disease and related dementias. On the basis of previous observations that tau is a direct substrate of histone deacetylase 6 (HDAC6), we sought to map all HDAC6-responsive sites in tau and determine how acetylation in a site-specific manner affects tau's biophysical properties in vitro Our findings indicate that several acetylation sites in tau are responsive to HDAC6 and that acetylation on Lys-321 (within a KCGS motif) is both essential for acetylation-mediated inhibition of tau aggregation in vitro and a molecular tactic for preventing phosphorylation on the downstream Ser-324 residue. To determine the functional consequence of this HDAC6-regulated phosphorylation event, we examined tau's ability to promote microtubule assembly and found that phosphorylation of Ser-324 interferes with the normal microtubule-stabilizing function of tau. Tau phosphorylation of Ser-324 (pSer-324) has not previously been evaluated in the context of tauopathy, and here we observed increased deposition of pSer-324-positive tau both in mouse models of tauopathy and in patients with Alzheimer's disease. These findings uncover a novel acetylation-phosphorylation switch at Lys-321/Ser-324 that coordinately regulates tau polymerization and function. Because the disease relevance of this finding is evident, additional studies are needed to examine the role of pSer-324 in tau pathobiology and to determine whether therapeutically modulating this acetylation-phosphorylation switch affects disease progression in vivo.

New self-management technologies for the treatment of hypertension: general practitioners' perspectives.

Background: Digital health interventions, such as those that can be delivered via smartphone applications (apps) or wireless blood pressure monitors, offer a new, scalable and potentially cost-effective way to improve hypertension self-management. In Ireland, as is common in the UK, the majority of hypertension management occurs in general practice. Therefore, it is crucial to investigate how general practitioners (GPs) feel about and engage with the growth of these new methods of self-management of blood pressure. Aim: To explore GPs' perspectives of self-management technology to support medication adherence and blood pressure control in patients with hypertension. Design and setting: This was a qualitative interview study based in the West of Ireland. Ten GPs who were purposively sampled participated in semi-structured interviews. Thematic analysis was carried out on the data. Results: Three major themes were identified: current reach and future potential, empowerment and responsibility. Conclusions: GPs could see the benefit of using these technologies, such as more accurate blood pressure data and potential to engage patients in self-management. Concerns relating to the increased workload associated with a potentially unmanageable quantity of information and an increase in healthcare use among the 'worried well' also emerged strongly from the data.

Implementation of the SMART MOVE intervention in primary care: a qualitative study using normalisation process theory.

BACKGROUND: Problematic translational gaps continue to exist between demonstrating the positive impact of healthcare interventions in research settings and their implementation into routine daily practice. The aim of this qualitative evaluation of the SMART MOVE trial was to conduct a theoretically informed analysis, using normalisation process theory, of the potential barriers and levers to the implementation of a mhealth intervention to promote physical activity in primary care. METHODS: The study took place in the West of Ireland with recruitment in the community from the Clare Primary Care Network. SMART MOVE trial participants and the staff from four primary care centres were invited to take part and all agreed to do so. A qualitative methodology with a combination of focus groups (general practitioners, practice nurses and non-clinical staff from four separate primary care centres, n = 14) and individual semi-structured interviews (intervention and control SMART MOVE trial participants, n = 4) with purposeful sampling utilising the principles of Framework Analysis was utilised. The Normalisation Process Theory was used to develop the topic guide for the interviews and also informed the data analysis process. RESULTS: Four themes emerged from the analysis: personal and professional exercise strategies; roles and responsibilities to support active engagement; utilisation challenges; and evaluation, adoption and adherence. It was evident that introducing a new healthcare intervention demands a comprehensive evaluation of the intervention itself and also the environment in which it is to operate. Despite certain obstacles, the opportunity exists for the successful implementation of a novel healthcare intervention that addresses a hitherto unresolved healthcare need, provided that the intervention has strong usability attributes for both disseminators and target users and coheres strongly with the core objectives and culture of the health care environment in which it is to operate. CONCLUSION: We carried out a theoretical analysis of stakeholder informed barriers and levers to the implementation of a novel exercise promotion tool in the Irish primary care setting. We believe that this process amplifies the implementation potential of such an intervention in primary care. The SMART MOVE trial is registered at Current Controlled Trials (ISRCTN99944116; Date of registration: 1st August 2012).

Tau deposition drives neuropathological, inflammatory and behavioral abnormalities independently of neuronal loss in a novel mouse model.

Aberrant tau protein accumulation drives neurofibrillary tangle (NFT) formation in several neurodegenerative diseases. Currently, efforts to elucidate pathogenic mechanisms and assess the efficacy of therapeutic targets are limited by constraints of existing models of tauopathy. In order to generate a more versatile mouse model of tauopathy, somatic brain transgenesis was utilized to deliver adeno-associated virus serotype 1 (AAV1) encoding human mutant P301L-tau compared with GFP control. At 6 months of age, we observed widespread human tau expression with concomitant accumulation of hyperphosphorylated and abnormally folded proteinase K resistant tau. However, no overt neuronal loss was observed, though significant abnormalities were noted in the postsynaptic scaffolding protein PSD95. Neurofibrillary pathology was also detected with Gallyas silver stain and Thioflavin-S, and electron microscopy revealed the deposition of closely packed filaments. In addition to classic markers of tauopathy, significant neuroinflammation and extensive gliosis were detected in AAV1-Tau(P301L) mice. This model also recapitulates the behavioral phenotype characteristic of mouse models of tauopathy, including abnormalities in exploration, anxiety, and learning and memory. These findings indicate that biochemical and neuropathological hallmarks of tauopathies are accurately conserved and are independent of cell death in this novel AAV-based model of tauopathy, which offers exceptional versatility and speed in comparison with existing transgenic models. Therefore, we anticipate this approach will facilitate the identification and validation of genetic modifiers of disease, as well as accelerate preclinical assessment of potential therapeutic targets.

(Patho-)physiological relevance of PINK1-dependent ubiquitin phosphorylation.

Mutations in PINK1 and PARKIN cause recessive, early-onset Parkinson's disease (PD). Together, these two proteins orchestrate a protective mitophagic response that ensures the safe disposal of damaged mitochondria. The kinase PINK1 phosphorylates ubiquitin (Ub) at the conserved residue S65, in addition to modifying the E3 ubiquitin ligase Parkin. The structural and functional consequences of Ub phosphorylation (pS65-Ub) have already been suggested from in vitro experiments, but its (patho-)physiological significance remains unknown. We have generated novel antibodies and assessed pS65-Ub signals in vitro and in cells, including primary neurons, under endogenous conditions. pS65-Ub is dependent on PINK1 kinase activity as confirmed in patient fibroblasts and postmortem brain samples harboring pathogenic mutations. We show that pS65-Ub is reversible and barely detectable under basal conditions, but rapidly induced upon mitochondrial stress in cells and amplified in the presence of functional Parkin. pS65-Ub accumulates in human brain during aging and disease in the form of cytoplasmic granules that partially overlap with mitochondrial, lysosomal, and total Ub markers. Additional studies are now warranted to further elucidate pS65-Ub functions and fully explore its potential for biomarker or therapeutic development.

A truncating SOD1 mutation, p.Gly141X, is associated with clinical and pathologic heterogeneity, including frontotemporal lobar degeneration.

Amyotrophic lateral sclerosis (ALS) is a degenerative disorder affecting upper and lower motor neurons, but it is increasingly recognized to affect other systems, with cognitive impairment resembling frontotemporal dementia (FTD) in some patients. We report clinical and pathologic findings of a family with ALS due to a truncating mutation, p.Gly141X, in copper/zinc superoxide dismutase (SOD1). The proband presented clinically with FTD and later showed progressive motor neuron disease, while all other family members had early-onset and rapidly progressive ALS without significant cognitive deficits. Pathologic examination of both the proband and her daughter revealed degeneration of corticospinal tracts and motor neurons in brain and spinal cord compatible with ALS. On the other hand, the proband also had neocortical and limbic system degeneration with pleomorphic neuronal cytoplasmic inclusions. Extramotor pathology in her daughter was relatively restricted to the hypothalamus and extrapyramidal system, but not the neocortex. The inclusions in the proband and her daughter were immunoreactive for SOD1, but negative for TAR DNA-binding protein of 43 kDa (TDP-43). In the proband, a number of the neocortical inclusions were immunopositive for α-internexin, initially suggesting a diagnosis of atypical FTLD, but there was no evidence of fused in sarcoma (FUS) immunoreactivity, which is often detected in atypical FTLD. Analogous to atypical FTLD, neuronal inclusions had variable co-localization of SOD1 and α-internexin. The current classification of FTLD is based on the major constituent protein: FTLD-tau, FTLD-TDP-43, and FTLD-FUS. The proband in this family indicates that SOD1, while rare, can also be the substrate of FTLD, in addition to the more common presentation of ALS. The explanation for clinical and pathologic heterogeneity of SOD1 mutations, including the p.Gly141X mutation, remains unresolved.