"Functional electrical stimulation (FES) impacted on important aspects of my life" - A qualitative exploration of chronic stroke patients' and carers' perceptions of FES in the management of dropped foot.

Our purpose was to explore the impact of functional electrical stimulation (FES), as applied in the management of dropped foot, on patients with chronic stroke and their carers. A qualitative phenomenological study was undertaken. Participants were recruited from the Lothian FES clinic. Data were collected by using semistructured face-to-face interviews and analysed by using the framework of interpretative phenomenological analysis (IPA). Thirteen patients and nine carers consented to participate in the study. The overarching theme that emerged from the data was that "FES impacted on important aspects of my life." Four subthemes were identified: 1) "Walking with FES is much better"; 2) "FES helped regain control of life"; 3) "Feeling good comes with using FES"; and 4) "FES is not perfect but it is of value." This study provides insights into the perceptions of patients and carers of the impact of FES. These results have implications for clinicians delivering an FES service for the management of dropped foot after stroke. © Informa Healthcare. USA, Inc

Carer experiences of life after stroke - a qualitative analysis

Aims and objectives. Carers' experiences of caring for a stroke survivor were explored, including reactions and changes in their lives. Method. A phenomenological approach was taken to the collection and analysis of data. Semi-structured interviews lasting an average of 43 min were carried out with nine informal carers in their own homes. All were married to someone who had survived a stroke. Results. An overarching theme emerged, entitled: 'lives turned upside-down'. It took time for participants to understand the long-term impacts of stroke. Carers experienced increased caring and domestic workloads alongside reduced participation and altered expectations of life. They found emotional and cognitive changes in their partners particularly distressing, and would have valued more information and help with adjusting to the increased emotional, physical and cognitive workload of caring. Conclusions. It is important to support carers of people who have survived a stroke in adjusting to their changed lifestyles. This may affect their quality of life as well as sustainability of caring, and requires further research

User experiences, preferences and choices relating to functional electrical stimulation and ankle foot orthoses for foot-drop after stroke

Objectives: To explore experiences, preferences and choices relating to the use of ankle foot orthoses (AFOs) and functional electrical stimulation (FES) for foot-drop by people who have suffered a stroke and their carers, with the aim of informing clinical decision-making. Design: Semi-structured interviews explored individual experiences through a phenomenological approach. The Interpretative Phenomenological Analysis framework was used to enable organisation and interpretation of qualitative interview data. Setting: Participants who had used both transcutaneous FES and one of several types of AFO were recruited from a single FES clinic. Participants: Nine people who had suffered a stroke and four carers were recruited purposively, including people between 2 and 9 years post stroke, with different degrees of difficulty in walking. Results: Participants described experiences, preferences and choices relating to AFO and FES use. All but one person expressed a preference for FES use and related this to being able to move the ankle more freely; walk more normally, safely and independently; and greater comfort. Several people also used AFOs when the FES equipment failed, when travelling and near water. One person rationed their use of FES on a daily basis due to allergic reactions. Conclusions: The consensus in this sample demonstrated positive and negative experiences of both FES and AFO use. Participants weighed up the pros and cons, and despite predominant preferences for FES, many also used AFOs due to some drawbacks of FES. Further research and development are required to reduce drawbacks and further explore users' experiences. © 2010 Chartered Society of Physiotherapy

Characterization of HSP90 isoforms in transformed bovine leukocytes infected with Theileria annulata

HSP90 chaperones are essential regulators of cellular function, as they ensure the appropriate conformation of multiple key client proteins. Four HSP90 isoforms were identified in the protozoan parasite Theileria annulata. Partial characterisation was undertaken for three and localisation confirmed for cytoplasmic (TA12105); endoplasmic reticulum (TA06470) and apicoplast (TA10720) forms. ATPase activity and binding to the HSP90 inhibitor geldanamycin, were demonstrated for recombinant TA12105 and all three native forms could be isolated to varying extents by binding to geldanamycin beads. Because it is essential, HSP90 is considered a potential therapeutic drug target. Resistance to the only specific Theileriacidal drug is increasing and one challenge for design of drugs that target the parasite is to limit the effect on the host. An in vitro cell culture system that allows comparison between uninfected bovine cells and the T. annulata-infected counterpart was utilised to test the effects of geldanamycin and the derivative 17-AAG. T. annulata-infected cells had greater tolerance to geldanamycin than uninfected cells yet exhibited significantly more sensitivity to 17-AAG. These findings suggest that parasite HSP90 isoform (s) can alter the drug sensitivity of infected host cells and that members of the Theileria HSP90 family are potential targets worthy of further investigation

A mixed methods service evaluation of a pilot functional electrical stimulation clinic for the correction of dropped foot in patients with chronic stroke.

Aim To undertake a service evaluation of the pilot Lothian functional electrical stimulation (FES) clinic using both quantitative and qualitative methods and clinical practice reflection.\ud Background Clinical guidelines recommend that FES, for the management of dropped foot after stroke, is delivered by a specialist team. However, little detail is provided about the structure and composition of the specialist team or model of service delivery. A pilot Lothian FES clinic was developed to explore the clinical value of providing such a service to stroke patients with dropped foot and identify any service modifications.\ud Methods Mixed methods were used to evaluate the service and included quantitative, qualitative and reflective components. Phase 1: Before and after service evaluation of patients attending the FES clinic between 2003 and 2007. Outcomes of gait velocity and cadence were recorded at initial clinic appointment and 6 months after application of FES. Phase 2: Qualitative research exploring patients with stroke and carers’ experiences of the FES clinic. Data were collected via semi-structured interviews. Phase 3: A reflection on the service delivery model. Participants: Phase 1: 40 consecutive out-patients with stroke; Phase 2: 13 out-patients with stroke and 9 carers; Phase 3: Three specialist physiotherapists engaged in running the FES clinic.\ud Findings Statistically significant improvements (p < 0.001) were demonstrated in gait velocity and cadence. Qualitatively, one super-ordinate theme ‘The FES clinic met my needs’ emerged. Within this were four sub-themes, namely 1. ‘Getting to grips with FES wasn’t difficult’; 2. ‘It’s great to know they’re there’; 3. ‘Meeting up with others really helps’ and 4. ‘The service is great but could be better’. On reflection, minor modifications were made to the service delivery model but overall the service met user needs. This dedicated FES clinic produced positive physical outcomes and met the needs of this chronic stroke population

Characterisation of infection associated microRNA and protein cargo in extracellular vesicles of Theileria annulata infected leukocytes

The protozoan parasites, Theileria annulata and T. parva are unique amongst intracellular eukaryotic pathogens as they induce a transformation‐like phenotype in their bovine host cell. T. annulata causes tropical theileriosis, which is frequently fatal, with infected leukocytes becoming metastatic and forming foci in multiple organs resulting in destruction of the lymphoid system. Exosomes, a sub‐set of extracellular vesicles (EV), are critical in metastatic progression in many cancers. Here we characterised the cargo of EV from a control bovine lymphosarcoma cell line (BL20) and BL20 infected with T. annulata (TBL20) by comparative mass spectrometry and miRNA profiling (data available via ProteomeXchange, identifier PXD010713 and NCBI GEO, accession number GSE118456, respectively). Ingenuity Pathway Analysis that many infection‐associated proteins essential to migration and extracellular matrix digestion were upregulated in EV from TBL20 cells compared to BL20 controls. An altered repertoire of host miRNA, many with known roles in tumor and/or infection biology was also observed. Focusing on the tumor suppressor miRNA, bta‐miR‐181a and bta‐miR‐181b, we identified putative mRNA targets and confirmed the interaction of bta‐miR181a with icam‐1. We propose that EV and their miRNA cargo play an important role in the manipulation of the host cell phenotype and the pathobiology of Theileria infection

Susceptibility to disease (tropical theileriosis) is associated with differential expression of host genes that possess motifs recognised by a pathogen DNA binding protein

BACKGROUND: Knowledge of factors that influence the outcome of infection are crucial for determining the risk of severe disease and requires the characterisation of pathogen-host interactions that have evolved to confer variable susceptibility to infection. Cattle infected by Theileria annulata show a wide range in disease severity. Native (Bos indicus) Sahiwal cattle are tolerant to infection, whereas exotic (Bos taurus) Holstein cattle are susceptible to acute disease. METHODOLOGY/PRINCIPAL FINDINGS: We used RNA-seq to assess whether Theileria infected cell lines from Sahiwal cattle display a different transcriptome profile compared to Holstein and screened for altered expression of parasite factors that could generate differences in host cell gene expression. Significant differences (<0.1 FDR) in the expression level of a large number (2211) of bovine genes were identified, with enrichment of genes associated with Type I IFN, cholesterol biosynthesis, oncogenesis and parasite infection. A screen for parasite factors found limited evidence for differential expression. However, the number and location of DNA motifs bound by the TashAT2 factor (TA20095) were found to differ between the genomes of B. indicus vs. B. taurus, and divergent motif patterns were identified in infection-associated genes differentially expressed between Sahiwal and Holstein infected cells. CONCLUSIONS/SIGNIFICANCE: We conclude that divergent pathogen-host molecular interactions that influence chromatin architecture of the infected cell are a major determinant in the generation of gene expression differences linked to disease susceptibility

A bovine lymphosarcoma cell line infected with theileria annulata exhibits an irreversible reconfiguration of host cell gene expression

Theileria annulata, an intracellular parasite of bovine lymphoid cells, induces substantial phenotypic alterations to its host cell including continuous proliferation, cytoskeletal changes and resistance to apoptosis. While parasite induced modulation of host cell signal transduction pathways and NFκB activation are established, there remains considerable speculation on the complexities of the parasite directed control mechanisms that govern these radical changes to the host cell. Our objectives in this study were to provide a comprehensive analysis of the global changes to host cell gene expression with emphasis on those that result from direct intervention by the parasite. By using comparative microarray analysis of an uninfected bovine cell line and its Theileria infected counterpart, in conjunction with use of the specific parasitacidal agent, buparvaquone, we have identified a large number of host cell gene expression changes that result from parasite infection. Our results indicate that the viable parasite can irreversibly modify the transformed phenotype of a bovine cell line. Fifty percent of genes with altered expression failed to show a reversible response to parasite death, a possible contributing factor to initiation of host cell apoptosis. The genes that did show an early predicted response to loss of parasite viability highlighted a sub-group of genes that are likely to be under direct control by parasite infection. Network and pathway analysis demonstrated that this sub-group is significantly enriched for genes involved in regulation of chromatin modification and gene expression. The results provide evidence that the Theileria parasite has the regulatory capacity to generate widespread change to host cell gene expression in a complex and largely irreversible manner

Theileria lestoquardi displays reduced genetic diversity relative to sympatric Theileria annulata in Oman

The Apicomplexan parasites, Theileria lestoquardi and Theileria annulata, the causative agents of theileriosis in small and large ruminants, are widespread in Oman, in areas where cattle, sheep and goats co-graze. Genetic analysis can provide insight into the dynamics of the parasite and the evolutionary relationship between species. Here we identified ten genetic markers (micro- and mini-satellites) spread across the T. lestoquardi genome, and confirmed their species specificity. We then genotyped T. lestoquardi in different regions in Oman. The genetic structures of T. lestoquardi populations were then compared with previously published data, for comparable panels of markers, for sympatric T. annulata isolates. In addition, we examined two antigen genes in T. annulata (Tams1 and Ta9) and their orthologues in T. lestoquardi (Tlms1 and Tl9). The genetic diversity and multiplicity of infection (MOI) were lower in T. lestoquardi (He = 0.64–0.77) than T. annulata (He = 0.83–0.85) in all populations. Very limited genetic differentiation was found among T. lestoquardi and T. annulata populations. In contrast, limited but significant linkage disequilibrium was observed within regional populations of each species. We identified eight T. annulata isolates in small ruminants; the diversity and MOI were lower among ovine/caprine compared to bovine. Sequence diversity of the antigen genes, Tams1 and Ta9 in T. annulata (π = 0.0733 and π = 0.155 respectively), was 10-fold and 3-fold higher than the orthologous Tlms1 and Tl9 in T. lestoquardi (π = 0.006 and π = 0.055, respectively). Despite a comparably high prevalence, T. lestoquardi has lower genetic diversity compared to sympatric T. annulata populations. There was no evidence of differentiation among populations of either species. In comparison to T. lestoquardi, T. annulata has a larger effective population size. While genetic exchange and recombination occur in both parasite species, the extent of diversity, overall, is less for T. lestoquardi. It is, therefore, likely that T. lestoquardi evolved from an ancestor of present day T. annulata and that this occurred either once or on a limited number of occasions