Bromodeoxyuridine induces keratin protein synthesis at a posttranscriptional level in human lung tumour cell lines

Keratin intermediate filaments are formed in epithelial cells in a cell- and tissue-specific manner, but much remains unknown regarding the mechanisms which control the synthesis of these proteins. We examined the effect of the differentiation modulation agent, bromodeoxyuridine (BrdU), on two human keratin-negative (by immunocytochemistry) lung cell lines, DLKP and H82, and showed immunohistochemically that treatment with 10 µM BrdU over 7 days induced K8 and K18 protein synthesis in both lines. Immunoprecipitation and Western blot analyses revealed low levels of K8 and K18 proteins in untreated cell homogenates. These levels increased following treatment with BrdU for 7 days. K8 and K18 mRNAs were detected by Northern blot and reverse transcriptase polymerase chain reaction analyses in both lines before BrdU treatment, but no increase in mRNA levels was observed in either cell line over 21 days of treatment. This suggests, firstly, that keratin synthesis is normally blocked at a posttranscriptional level in DLKP and H82 cells, and secondly, that BrdU can reverse this block. A549 is a human lung cell line which contains K8 and K18 proteins. Treatment with BrdU increased K8 and K18 protein levels in these cells. No corresponding increase in K8 mRNA levels occurred, while an apparent increase in K18 mRNA levels was detected. HL-60 is a leukaemic cell line of haematopoietic rather than epithelial lineage which contains K8 and K18 mRNA transcripts prior to BrdU treatment, but does not contain keratin proteins. Again, K8 and K18 mRNA levels remained unchanged during BrdU treatment. However, neither K8 nor K18 proteins were detected following treatment, although BrdU is known to alter expression of other genes in HL-60 cells. BrdU thus appears to act at a posttranscriptional level and in an epithelial-specific manner to reverse a block in keratin synthesis in keratinnegative lung cancer cells and increase synthesis in keratin-positive lung cancer cells. This may represent a regulatory step in early lung development or a mechanism whereby tumour cells downregulate expression of a differentiated phenotype

Bromodeoxyuridine induces keratin protein synthesis at a posttranscriptional level in human lung tumour cell lines

Keratin intermediate filaments are formed in epithelial cells in a cell- and tissue-specific manner, but much remains unknown regarding the mechanisms which control the synthesis of these proteins. We examined the effect of the differentiation modulation agent, bromodeoxyuridine (BrdU), on two human keratin-negative (by immunocytochemistry) lung cell lines, DLKP and H82, and showed immunohistochemically that treatment with 10 µM BrdU over 7 days induced K8 and K18 protein synthesis in both lines. Immunoprecipitation and Western blot analyses revealed low levels of K8 and K18 proteins in untreated cell homogenates. These levels increased following treatment with BrdU for 7 days. K8 and K18 mRNAs were detected by Northern blot and reverse transcriptase polymerase chain reaction analyses in both lines before BrdU treatment, but no increase in mRNA levels was observed in either cell line over 21 days of treatment. This suggests, firstly, that keratin synthesis is normally blocked at a posttranscriptional level in DLKP and H82 cells, and secondly, that BrdU can reverse this block. A549 is a human lung cell line which contains K8 and K18 proteins. Treatment with BrdU increased K8 and K18 protein levels in these cells. No corresponding increase in K8 mRNA levels occurred, while an apparent increase in K18 mRNA levels was detected. HL-60 is a leukaemic cell line of haematopoietic rather than epithelial lineage which contains K8 and K18 mRNA transcripts prior to BrdU treatment, but does not contain keratin proteins. Again, K8 and K18 mRNA levels remained unchanged during BrdU treatment. However, neither K8 nor K18 proteins were detected following treatment, although BrdU is known to alter expression of other genes in HL-60 cells. BrdU thus appears to act at a posttranscriptional level and in an epithelial-specific manner to reverse a block in keratin synthesis in keratinnegative lung cancer cells and increase synthesis in keratin-positive lung cancer cells. This may represent a regulatory step in early lung development or a mechanism whereby tumour cells downregulate expression of a differentiated phenotype

Technology use among patients with cardiovascular disease: an assessment of patient need for a technology enabled behavioural change intervention.

Effective Cardiac Rehabilitation (CR) can significantly improve mortality and morbidity rates in relation to cardiovascular disease; however, uptake of traditional community-based long-term is very low. PATHway (Physical Activity Towards Health) will provide individualized rehabilitation programs, through an internet-enabled sensor-based home exercise platform that allows remote participation. The purpose of this study was to assess the level of interest and use of technology by individuals living with CVD in order to inform the design of a technology-enabled CR programme. Method: A technology usage questionnaire based on a previous study investigating the role of technology and mHealth in a CVD population was used (Dale et al., 2014) to ascertain the current level of technology use. All patients attending the Phase Four community cardiac rehabilitation HeartSmart programme (MedEx) were recruited (N=67; 66.2 years, SD= 8.55, Males =76.1%, Females=20.9%). Results: Technology usage was high with 60% of participants owning a smartphone and 85% accessing the internet (54% of whom access it everyday). Participants endorsed the idea of technology enabled cardiac rehabilitation, indicating that they found the idea ‘ appealing’. 79% were interested in receiving ongoing CR support via their smartphones, 79% were interested in receiving CR via the internet. It was found that 52% of patients found the idea of a virtual rehabilitation class appealing. Conclusion: This study provides support for the patient need for a technology enabled behavioural change intervention, specifically through the provision of an internet-enabled sensor-based home exercise platform that allows remote participation in CR exercise programs

Uptake to a community based chronic illness rehabilitation programme (CBCIR): Is there a gender disparity?

Background: Multi-morbidity and chronic conditions pose a threat to population health. Despite known benefits of rehabilitation using structured exercise, uptake to such programmes remain sub-optimal. The aim of this study is to identify the psychosocial and health related fitness correlates of uptake to a CBCIR in men and women, with the secondary aim of identifying the rate of uptake. Methods: Participants referred to a CBCIR via GPs and hospitals completed an induction process. This introduced them to the CBCIR programme and got them to complete a multi-section questionnaire (including instruments on physical activity, exercise self-efficacy, intentions for exercise, and perceived family/friend social support) and complete a battery of physical health measures (including the Incremental shuttle walk test (ISWT), a lower body strength test and body mass index (BMI)). Post induction participants who attended an exercise class were classified as ‘Uptakers’, whilst those who never came back were classified as ‘Non-Uptakers’. Class attendance was objectively monitored by the researchers. Data were analysed using SPSS, and are presented using means, standard deviations and proportions, group differences are examined via t-tests and logistic regression was used to predict uptake. Results: A total of 441 participants (56% male; average age 64.3 ±12 years completed induction measures. Overall, 77% were identified as Uptakers (81% female, 74% male, p=0.068, 2-sided). Among men, Uptakers reported more days of 30mins moderate to vigorous physical activity (t(111) = -2.499,

Co-design and user validation of the MedFit App: a focus group analysis

This abstract details the development phase of the formative research process outlined by the Medical Research Council, used to develop a theoretically informed Android App, named MedFit, to enhance disease self-management and quality of life in adults with cardiovascular disease (CVD). The overall aim of the app is to increase physical activity minutes of adults with CVD. A key part of the development phase, which is the focus of this abstract, is the co-design and user validation of the MedFit app

Behavior Change Techniques in Physical Activity eHealth Interventions for People With Cardiovascular Disease: Systematic Review

Background: Cardiovascular disease (CVD) is the leading cause of premature death and disability in Europe, accounting for 4 million deaths per year and costing the European Union economy almost €196 billion annually. There is strong evidence to suggest that exercise-based secondary rehabilitation programs can decrease the mortality risk and improve health among patients with CVD. Theory-informed use of behavior change techniques (BCTs) is important in the design of cardiac rehabilitation programs aimed at changing cardiovascular risk factors. Electronic health (eHealth) is the use of information and communication technologies (ICTs) for health. This emerging area of health care has the ability to enhance self-management of chronic disease by making health care more accessible, affordable, and available to the public. However, evidence-based information on the use of BCTs in eHealth interventions is limited, and particularly so, for individuals living with CVD. Objective: The aim of this systematic review was to assess the application of BCTs in eHealth interventions designed to increase physical activity (PA) in CVD populations. Methods: A total of 7 electronic databases, including EBSCOhost (MEDLINE, PsycINFO, Academic Search Complete, SPORTDiscus with Full Text, and CINAHL Complete), Scopus, and Web of Science (Core Collection) were searched. Two authors independently reviewed references using the software package Covidence (Veritas Health Innovation). The reviewers met to resolve any discrepancies, with a third independent reviewer acting as an arbitrator when required. Following this, data were extracted from the papers that met the inclusion criteria. Bias assessment of the studies was carried out using the Cochrane Collaboration’s tool for assessing the risk of bias within Covidence; this was followed by a narrative synthesis. Results: Out of the 987 studies that were identified, 14 were included in the review. An additional 9 studies were added following a hand search of review paper references. The average number of BCTs used across the 23 studies was 7.2 (range 1-19). The top three most frequently used BCTs included information about health consequences (78%, 18/23), goal setting (behavior; 74%, 17/23), and joint third, self-monitoring of behavior and social support (practical) were included in 11 studies (48%, 11/23) each. Conclusions: This systematic review is the first to investigate the use of BCTs in PA eHealth interventions specifically designed for people with CVD. This research will have clear implications for health care policy and research by outlining the BCTs used in eHealth interventions for chronic illnesses, in particular CVD, thereby providing clear foundations for further research and developments in the area

MedFit: The development of a mobile-application to enhance participant self-management of their cardiovascular disease

Background: Cardiovascular disease (CVD) is the leading cause of premature death and disability in Europe. Cardiac Rehabilitation (CR) can reduce the impact of CVD by lowering mortality and morbidity rates and promoting healthy active lifestyles. Yet adherence within CR is low. Research suggests that mHealth interventions are useful in supporting the self-management of chronic disease. The purpose of this research is to report on the development of an mHealth intervention. Methods: For the intervention development the Medical Research Council’s formative process consisting of 4 stages; i) development, ii) feasibility/piloting, iii) evaluation and iv) implementation will be used to develop a theoretically informed Android App to enhance disease self-management and quality of life in CVD. Like CR it will use exercise as its main modality, and provide advice on other health behaviours. Results: A systematic review of the use of behaviour change techniques (BCTs) in physical activity eHealth interventions for CVD patients has been conducted. Seven electronic databases yielded 987 articles, 97 of which met the inclusion criteria for full text review. A multidisciplinary team comprised of exercise scientists, health behaviour change and technology specialists are using this information to develop the intervention prototype. Stage one will be followed by qualitative research, where end-users will be asked to examine the intervention in order to determine its feasibility and acceptability, to ultimately improve its efficacy through a co-design process. Conclusion: Preliminary findings and systematic review protocol will be reported as per the PRISMA guidelines, ultimately aiding the development of the MedFit app

A between sex analysis of self and proxy efficacy and its relationship with attendance at a community based chronic illness rehabilitation programme.

Background: Physical activity is a principal intervention in primary and secondary prevention of chronic illness. While the benefits of community based rehabilitation is acknowledged,the rate of attendance and adherence remains sub optimal. Self-efficacy is acknowledged to be a key factor relating to adherence in rehabilitation but the construct of proxy efficacy, defined as one’s confidence in the skills and abilities of a third party to function effectively on another’s behalf, is less often studied. This paper examines the relationship between gender and ‘self’ and ‘proxy-efficacy’ and their potential role in attendance to a community based chronic illness rehabiliation (CBCIR) programme. Methods: Participants attending induction at the programme completed a questionnaire assessing demographics and both self (9 items) and proxy efficacy for exercise (9 items) which was assessed on a Likert scale from 0 (not confident at all) to 10 (very confident), with a higher score indicating greater efficacy. Subsequently, attendance was objectively monitored by researchers at exercise sessions for 24 weeks. Results: 69 participants (M age=65.5 +9.8 years, 56% Male) completed all measures. Mean proxy-efficacy and mean self-efficacy is significantly greater in women compared to men (p<0.05). Mean proxy-efficacy was found to be higher than mean self-efficacy in both males (proxy-efficacy=8.7+1.2, self efficacy=8.3+1.8) and females (proxyefficacy= 7.7+2.2, self-efficacy=6.5+2.0) with no correlation between the variables. Correlational analyses found a moderate significant positive correlation between proxy efficacy and number of weeks attended in the 24 weeks (r=.440,p<.05) in women with no correlation observed between self-efficacy and weeks attended. No correlations are observed between either form of efficacy and attendance in men. Discussion: Participants report greater confidence in the proxy agent than themselves as the agent. This was furthered in women with an association identified between proxy efficacy and 24-week attendance. These results suggest the importance of the proxy for women in a CBCIR setting but further longitudinal research should be carried out in the area of proxy efficacy and attendance at community based chronic illness rehabilitation programmes. Implications: An understanding of these variables help CBCIR programme facilitators intervene to ensure greater attendance

T- and L-type Ca 2+ currents in freshly dispersed smooth muscle cells from the human proximal urethra

The purpose of the present study was to characterise Ca 2+ currents in smooth muscle cells solated from biopsy samples taken from the proximal urethra of patients undergoing surgery for bladder or prostate cancer. Cells were studied at 37°C using the amphotericin B perforated-patch onfiguration of the patch-clamp technique. Currents were recorded using Cs + -rich pipette solutions to block K + currents. Two components of current, with electrophysiological and pharmacological properties typical of T- and L-type Ca 2+ currents, were present in these cells. When steady-state inactivation curves for the L current were fitted with a Boltzmann equation, this yielded a VÎ of _45 ± 5 mV. In contrast, the T current inactivated with a VÎ of _80 ± 3 mV. The L currents were reduced in a concentration-dependent manner by nifedipine (ED50 = 159 ± 54 nM) and Ni 2+ (ED50 = 65 ± 16 mM) but were enhanced when external Ca 2+ was substituted with Ba 2+ . The T current was little affected by TTX, reduction in external Na + , application of nifedipine at concentrations below 300 nM or substitution of external Ca 2+ with Ba 2+ , but was reduced by Ni 2+ with an ED50 of 6 ± 1 mM. When cells were stepped from _100 to _30 mV in Ca 2+ -free conditions, small inward currents could be detected. These were enhanced 40-fold in divalent-cation-free solution and blocked in a concentration-dependent manner by Mg 2+ with an ED50 of 32 ± 16 mM. These data support the idea that human urethral myocytes possess currents with electrophysiological and pharmacological properties typical of T- and L-type Ca 2+ currents

MedFit: a mobile application for recovering CVD patients

The third phase of the recovery from cardiovascular disease (CVD) is an exercise-based rehabilitation programme. However, adherence to an exercise regime is typically not maintained by the patient for a variety of reasons such as lack of time, financial constraints, etc. In order to facilitate patients to perform their exercises from the comfort of their home and at their own convenience, we have developed a mobile application, termed MedFit. It provides access to a tailored suite of exercises along with easy to understand guidance from audio and video instructions. Two types of wearable sensors are utilized to allow motivational feedback to be provided to the user for self monitoring and to provide near real-time feedback. Fitbit, a commercially available activity and fitness tracker, is used to provide in-depth feedback for self-monitoring over longer periods of time (e.g. day, week, month), whereas the Shimmer wireless sensing platform provides the data for near real-time feedback on the quality of the exercises performed. MedFit is a simple and intuitive mobile application designed to provide the motivation and tools for patients to help ensure faster recovery from the trauma caused by CVD. In this paper we describe the MedFit application as a demo submission to the 2nd MMHealth Workshop at ACM MM 2017