Reference values for wrist-worn accelerometer physical activity metrics in England children and adolescents.

BACKGROUND: Over the last decade use of raw acceleration metrics to assess physical activity has increased. Metrics such as Euclidean Norm Minus One (ENMO), and Mean Amplitude Deviation (MAD) can be used to generate metrics which describe physical activity volume (average acceleration), intensity distribution (intensity gradient), and intensity of the most active periods (MX metrics) of the day. Presently, relatively little comparative data for these metrics exists in youth. To address this need, this study presents age- and sex-specific reference percentile values in England youth and compares physical activity volume and intensity profiles by age and sex. METHODS: Wrist-worn accelerometer data from 10 studies involving youth aged 5 to 15 y were pooled. Weekday and weekend waking hours were first calculated for youth in school Years (Y) 1&2, Y4&5, Y6&7, and Y8&9 to determine waking hours durations by age-groups and day types. A valid waking hours day was defined as accelerometer wear for ≥ 600 min·d-1 and participants with ≥ 3 valid weekdays and ≥ 1 valid weekend day were included. Mean ENMO- and MAD-generated average acceleration, intensity gradient, and MX metrics were calculated and summarised as weighted week averages. Sex-specific smoothed percentile curves were generated for each metric using Generalized Additive Models for Location Scale and Shape. Linear mixed models examined age and sex differences. RESULTS: The analytical sample included 1250 participants. Physical activity peaked between ages 6.5-10.5 y, depending on metric. For all metrics the highest activity levels occurred in less active participants (3rd-50th percentile) and girls, 0.5 to 1.5 y earlier than more active peers, and boys, respectively. Irrespective of metric, boys were more active than girls (p < .001) and physical activity was lowest in the Y8&9 group, particularly when compared to the Y1&2 group (p < .001). CONCLUSIONS: Percentile reference values for average acceleration, intensity gradient, and MX metrics have utility in describing age- and sex-specific values for physical activity volume and intensity in youth. There is a need to generate nationally-representative wrist-acceleration population-referenced norms for these metrics to further facilitate health-related physical activity research and promotion.

The Physical Behaviour Intensity Spectrum and Body Mass Index in School-Aged Youth: A Compositional Analysis of Pooled Individual Participant Data.

We examined the compositional associations between the intensity spectrum derived from incremental acceleration intensity bands and the body mass index (BMI) z-score in youth, and investigated the estimated differences in BMI z-score following time reallocations between intensity bands. School-aged youth from 63 schools wore wrist accelerometers, and data of 1453 participants (57.5% girls) were analysed. Nine acceleration intensity bands (range: 0−50 mg to ≥700 mg) were used to generate time-use compositions. Multivariate regression assessed the associations between intensity band compositions and BMI z-scores. Compositional isotemporal substitution estimated the differences in BMI z-score following time reallocations between intensity bands. The ≥700 mg intensity bandwas strongly and inversely associated with BMI z-score (p < 0.001). The estimated differences in BMI z-score when 5 min were reallocated to and from the ≥700 mg band and reallocated equally among the remaining bands were −0.28 and 0.44, respectively (boys), and −0.39 and 1.06, respectively (girls). The time in the ≥700 mg intensity band was significantly associated with BMI z-score, irrespective of sex. When even modest durations of time in this band were reallocated, the asymmetrical estimated differences in BMI z-score were clinically meaningful. The findings highlight the utility of the full physical activity intensity spectrum over a priori-determined absolute intensity cut-point approaches.

Clinical exercise provision in the UK: comparison of staff job titles, roles and qualifications across five specialised exercise services.

OBJECTIVES: In the UK, the National Health Service long-term plan advocates exercise as a key component of clinical services, but there is no clearly defined workforce to deliver the plan. We aimed to provide an overview of current UK clinical exercise services, focusing on exercise staff job titles, roles and qualifications across cardiovascular, respiratory, stroke, falls and cancer services. METHODS: Clinical exercise services were identified electronically between May 2020 and September 2020 using publicly available information from clinical commissioning groups, national health boards and published audit data. Data relating to staff job titles, roles, qualifications and exercise delivery were collected via electronic records and telephone/email contact with service providers. RESULTS: Data were obtained for 731 of 890 eligible clinical services (216 cardiac, 162 respiratory, 129 stroke, 117 falls, 107 cancer). Cardiac rehabilitation services provided both clinical (phase III) and community (phase IV) exercise interventions delivered by physiotherapists, exercise physiologists (exercise specific BSc/MSc) and exercise instructors (vocationally qualified with or without BSc/MSc). Respiratory, stroke and falls services provided a clinical exercise intervention only, mostly delivered by physiotherapists and occupational therapists. Cancer services provided a community exercise service only, delivered by vocationally qualified exercise instructors. Job titles of 'exercise physiologists' (n=115) bore little alignment to their qualifications, with a large heterogeneity across services. CONCLUSION: In the UK, clinical exercise services job titles, roles and qualifications were inconsistent. Regulation of exercise job titles and roles is required to remove the current disparities in this area.

Charter to establish clinical exercise physiology as a recognised allied health profession in the UK: a call to action.

The UK population is growing, ageing and becoming increasingly inactive and unfit. Personalised and targeted exercise interventions are beneficial for ageing and the management of chronic and complex conditions. Increasing the uptake of effective exercise and physical activity (PA) interventions is vital to support a healthier society and decrease healthcare costs. Current strategies for exercise and PA at a population level mostly involve self-directed exercise pathways, delivered largely via the fitness industry. Even for those who opt-in and manage to achieve the current recommendations regarding minimum PA, this generic 'one-size-fits-all' approach often fails to demonstrate meaningful physiological and health benefits. Personalised exercise prescription and appropriate exercise testing, monitoring and progression of interventions for individuals with chronic disease should be provided by appropriately trained and recognised exercise healthcare professionals, educated in the cognate disciplines of exercise science (eg, physiology, biomechanics, motor control, psychology). This workforce has operated for >20 years in the Australian public and private healthcare systems. Accredited exercise physiologists (AEPs) are recognised allied health professionals, with demonstrable health and economic benefits. AEPs have knowledge of the risks and benefits of distinct forms of exercise, skills in the personalised prescription and optimal delivery of exercise, and competencies to support sustained PA behavioural change, based on the established scientific evidence. In this charter, we propose a road map for the training, accreditation and promotion of a clinical exercise physiology profession in the UK.

Sit Less and Move More-A Multicomponent Intervention With and Without Height-Adjustable Workstations in Contact Center Call Agents: A Pilot Randomized Controlled Trial.

OBJECTIVE: To pilot a multicomponent intervention to sit less and move more, with (SLAMM+) and without (SLAMM) height-adjustable workstations, in contact center call agents. METHODS: Agents were individually randomized to SLAMM or SLAMM+ in this 10-month, parallel, open-label, pilot trial. Mixed-methods assessed response, recruitment, retention, attrition and completion rates, adverse effects, trial feasibility and acceptability, preliminary effectiveness on worktime sitting, and described secondary outcomes. RESULTS: The participant recruitment rate, and randomization, data collection, and interventions were mostly acceptable. Refinements to organization recruitment were identified. High staff turnover negatively impacted retention and completion rates. The multicomponent intervention with height-adjustable workstations has potential to reduce sitting time at work. CONCLUSIONS: The demonstrated findings will help prepare for a future randomized controlled trial designed to assess the effect of the interventions.

Using an e-Health Intervention to Reduce Prolonged Sitting in UK Office Workers: A Randomised Acceptability and Feasibility Study.

Low-cost workplace interventions are required to reduce prolonged sitting in office workers as this may improve employees' health and well-being. This study aimed to assess the acceptability and feasibility of an e-health intervention to reduce prolonged sitting among sedentary UK-based office workers. Secondary aims were to describe preliminary changes in employee health, mood and work productivity after using an e-health intervention. Healthy, university office workers (n = 14) completed this study. An 8 week randomised crossover design was used, consisting of two trials: Intervention (computer-based prompts) and Control. Eligibility and retention rates were recorded to assess the feasibility of the trial and interviews were conducted following the intervention to explore its acceptability. Sitting, standing and stepping were objectively assessed prior to and during week 8 of each trial. Before and after each trial, measurements of vascular function, cerebrovascular function, mood and work productivity were obtained. This study had eligibility and retention rates of 54.5% and 77.8%, respectively. Participants expressed a lack of autonomy and disruption to their workflow when using the e-health intervention, raising concerns over its acceptability and long-term implementation. Preliminary data indicate that the intervention may improve the patterning of activity accrued during work hours, with increases in the number of standing and stepping bouts completed, in addition to improving vascular function. This e-health intervention is feasible to deliver in a cohort of university office workers. However, adaptations to its implementation, such as personalised settings, are needed to increase acceptability before larger trials can be conducted.

Validity and reliability of subjective methods to assess sedentary behaviour in adults: a systematic review and meta-analysis.

BACKGROUND: Subjective measures of sedentary behaviour (SB) (i.e. questionnaires and diaries/logs) are widely implemented, and can be useful for capturing type and context of SBs. However, little is known about comparative validity and reliability. The aim of this systematic review and meta-analysis was to: 1) identify subjective methods to assess overall, domain- and behaviour-specific SB, and 2) examine the validity and reliability of these methods. METHODS: The databases MEDLINE, EMBASE and SPORTDiscus were searched up to March 2020. Inclusion criteria were: 1) assessment of SB, 2) evaluation of subjective measurement tools, 3) being performed in healthy adults, 4) manuscript written in English, and 5) paper was peer-reviewed. Data of validity and/or reliability measurements was extracted from included studies and a meta-analysis using random effects was performed to assess the pooled correlation coefficients of the validity. RESULTS: The systematic search resulted in 2423 hits. After excluding duplicates and screening on title and abstract, 82 studies were included with 75 self-reported measurement tools. There was wide variability in the measurement properties and quality of the studies. The criterion validity varied between poor-to-excellent (correlation coefficient [R] range - 0.01- 0.90) with logs/diaries (R = 0.63 [95%CI 0.48-0.78]) showing higher criterion validity compared to questionnaires (R = 0.35 [95%CI 0.32-0.39]). Furthermore, correlation coefficients of single- and multiple-item questionnaires were comparable (1-item R = 0.34; 2-to-9-items R = 0.35; ≥10-items R = 0.37). The reliability of SB measures was moderate-to-good, with the quality of these studies being mostly fair-to-good. CONCLUSION: Logs and diaries are recommended to validly and reliably assess self-reported SB. However, due to time and resources constraints, 1-item questionnaires may be preferred to subjectively assess SB in large-scale observations when showing similar validity and reliability compared to longer questionnaires. REGISTRATION NUMBER: CRD42018105994.

Temporal dynamics of sitting behavior at work.

Sitting for prolonged periods of time impairs people's health. Prior research has mainly investigated sitting behavior on an aggregate level, for example, by analyzing total sitting time per day. By contrast, taking a dynamic approach, here we conceptualize sitting behavior as a continuous chain of sit-to-stand and stand-to-sit transitions. We use multilevel time-to-event analysis to analyze the timing of these transitions. We analyze ∼30,000 objectively measured posture transitions from 156 people during work time. Results indicate that the temporal dynamics of sit-to-stand transitions differ from stand-to-sit transitions, and that people are quicker to switch postures later in the workday, and quicker to stand up after having been more active in the recent hours. We found no evidence for associations with physical fitness. Altogether, these findings provide insights into the origins of people's stand-up and sit-down decisions, show that sitting behavior is fundamentally different from exercise behavior, and provide pointers for the development of interventions.

A multi-component intervention to sit less and move more in a contact centre setting: a feasibility study.

BACKGROUND: Call agents spend ~ 90% of their working day seated, which may negatively impact health, productivity, and wellbeing. This study aimed to explore the acceptability and feasibility of a multi-component workplace intervention targeting increased activity and decreased prolonged sitting in the contact centre setting prior to a full-scale effectiveness trial. METHODS: An 8-week non-randomised pre-post feasibility study was conducted. Using a mixed methods approach, focus groups and interviews were thematically analysed to explore the acceptability and feasibility of key study phases, and provide context to agents' process evaluation and survey responses. The multi-component intervention, conducted in a single call centre, included height-adjustable workstations, emails, education and training sessions, and support from team leaders and a workplace champion. RESULTS: Six (of 20) team leaders were recruited, with 17 of 84 call agents (78% female, 39.3 ± 11.9 years) completing baseline assessments and 13 completing follow-up. High workload influenced recruitment. Call agents perceived assessments as acceptable, though strategies are needed to enhance fidelity. Education sessions, height-adjustable workstations and emails were perceived as the most effective components; however, height-adjustable hot-desks were not perceived as feasible in this setting. CONCLUSIONS: This study has identified unique, pragmatic considerations for conducting a multi-level, multi-component PA and SB intervention and associated evaluation in highly sedentary call agents in the challenging contact centre setting. The intervention was largely perceived positively, with call agents and team leaders describing numerous perceived positive effects on behavioural, health and work-related outcomes. Findings will be of value to researchers attempting to intervene in contact centres and will be used by the current authors to design a subsequent trial.

Carotid Artery Function Is Restored in Subjects With Elevated Cardiovascular Disease Risk After a 12-Week Physical Activity Intervention.

Sympathetic nervous system activation elicits carotid artery vasodilation in healthy subjects, yet vasoconstriction in those with cardiovascular disease (CVD). Whether carotid artery vasoconstriction can be reversed is currently unknown. Nineteen subjects with increased risk for CVD were referred to a 12-week physical activity intervention, and 12 participants with increased risk for CVD were recruited as a no treatment control group. Cardiorespiratory and vascular health measures were collected at baseline and 12 weeks. Results indicate that carotid artery vasoconstriction in response to sympathetic stimulation may be reversed in subjects at increased risk of CVD. These findings warrant further investigation.

Educator perspectives on factors influencing children's school-based physical activity.

Formative research is an important first step in the design and development of children's school-based physical activity (PA) interventions. Exploration of educator [headteacher and physical education (PE)-co-ordinator] perceptions toward the promotion of school-based PA, including PE delivery has however been limited. This study took a socio-ecological approach to explore the barriers and facilitators of children's school-based PA from the perspective of school educators. Interviews were conducted with headteachers (n = 4), PE-co-ordinators (n = 4) and a deputy headteacher (n = 1) and data thematically analysed using Nvivo software (version 10). Findings suggested that, at an organizational level headteachers were the predominant driving force in the promotion of PA opportunities, yet institutional barriers including low priority for PA and PE were perceived to negate delivery. At an interpersonal level, strategies to increase the delivery of school-based PA were developed, however poor teacher-coach relationships and significant others reduced PA promotion opportunities. Child PA was further negated through intrapersonal factors, including lack of PE-specific teacher training and varying teacher interest in PA and sport. To increase primary school children's school-based PA, barriers and facilitators at the organizational, interpersonal and intrapersonal level must be considered and targeted and researchers and schools should work in partnership to develop future interventions.

A formative study exploring perceptions of physical activity and physical activity monitoring among children and young people with cystic fibrosis and health care professionals.

BACKGROUND: Physical activity (PA) is associated with reduced hospitalisations and maintenance of lung function in patients with Cystic Fibrosis (CF). PA is therefore recommended as part of standard care. Despite this, there is no consensus for monitoring of PA and little is known about perceptions of PA monitoring among children and young people with CF. Therefore, the research aimed to explore patients' perceptions of PA and the acceptability of using PA monitoring devices with children and young people with CF. METHODS: An action research approach was utilised, whereby findings from earlier research phases informed subsequent phases. Four phases were utilised, including patient interviews, PA monitoring, follow-up patient interviews and health care professional (HCP) interviews. Subsequently, an expert panel discussed the study to develop recommendations for practice and future research. RESULTS: Findings suggest that experiences of PA in children and young people with CF are largely comparable to their non-CF peers, with individuals engaging in a variety of activities. CF was not perceived as a barrier per se, although participants acknowledged that they could be limited by their symptoms. Maintenance of health emerged as a key facilitator, in some cases PA offered patients the opportunity to 'normalise' their condition. Participants reported enjoying wearing the monitoring devices and had good compliance. Wrist-worn devices and devices providing feedback were preferred. HCPs recognised the potential benefits of the devices in clinical practice. Recommendations based on these findings are that interventions to promote PA in children and young people with CF should be individualised and involve families to promote PA as part of an active lifestyle. Patients should receive support alongside the PA data obtained from monitoring devices. CONCLUSIONS: PA monitoring devices appear to be an acceptable method for objective assessment of PA among children and young people with CF and their clinicians. Wrist-worn devices, which are unobtrusive and can display feedback, were perceived as most acceptable. By understanding the factors impacting PA, CF health professionals will be better placed to support patients and improve health outcomes.

Individual calibration of accelerometers in children and their health-related implications.

This study compared children's physical activity (PA) levels, the prevalence of children meeting current guidelines of ≥60 minutes of daily moderate to vigorous PA (MVPA), and PA-health associations using individually calibrated (IC) and empirical accelerometer cutpoints. Data from 75 (n = 32 boys) 10-12 year old children were included in this study. Clustered cardiometabolic (CM) risk, directly measured cardiorespiratory fitness (CRF), anthropometric and 7 day accelerometer data were included within analysis. PA data were classified using Froude anchored IC, Evenson et al. (Evenson, K. R., Catellier, D. J., Gill, K., Ondrak, K. S., & McMurray, R. G. (2008). Calibration of two objective measures of physical activity for children. Journal of Sports Sciences, 26(14), 1557-1565. doi:10.1080/02640410802334196) (Ev) and Mackintosh et al. (Mackintosh, K. A., Fairclough, S. J., Stratton, G., & Ridgers, N. D. (2012). A calibration protocol for population-specific accelerometer cutpoints in children. PLoS One, 7(5), e36919. doi:10.1371/journal.pone.0036919) (Mack) cutpoints. The proportion of the cohort meeting ≥60mins MVPA/day ranged from 37%-56% depending on the cutpoints used. Reported PA differed significantly across the cutpoint sets. IC LPA and MPA were predictors of CRF (LPA: standardised ß = 0.32, p = 0.002, MPA: standardised ß = 0.27 p = 0.013). IC MPA also predicted BMI Z-score (standardised ß = -0.35, p = 0.004). Ev VPA was a predictor of BMI Z-score (standardised ß = -0.33, p = 0.012). Cutpoint choice has a substantial impact on reported PA levels though no significant associations with CM risk were observed. Froude IC cutpoints represent a promising approach towards classifying children's PA data.

From Surveillance to Intervention: Overview and Baseline Findings for the Active City of Liverpool Active Schools and SportsLinx (A-CLASS) Project.

This paper outlines the implementation of a programme of work that started with the development of a population-level children's health, fitness and lifestyle study in 1996 (SportsLinx) leading to selected interventions one of which is described in detail: the Active City of Liverpool, Active Schools and SportsLinx (A-CLASS) Project. The A-CLASS Project aimed to quantify the effectiveness of structured and unstructured physical activity (PA) programmes on children's PA, fitness, body composition, bone health, cardiac and vascular structures, fundamental movement skills, physical self-perception and self-esteem. The study was a four-arm parallel-group school-based cluster randomised controlled trial (clinical trials no. NCT02963805), and compared different exposure groups: a high intensity PA (HIPA) group, a fundamental movement skill (FMS) group, a PA signposting (PASS) group and a control group, in a two-schools-per-condition design. Baseline findings indicate that children's fundamental movement skill competence levels are low-to-moderate, yet these skills are inversely associated with percentage body fat. Outcomes of this project will make an important contribution to the design and implementation of children's PA promotion initiatives.

Utility of three anthropometric indices in assessing the cardiometabolic risk profile in children.

OBJECTIVES: To evaluate the ability of BMI, WC, and WHtR to identify increased cardiometabolic risk in pre-adolescents. METHODS: This is a cross-sectional study involving 192 children (10.92 ± 0.58 years, 56% female) from the United Kingdom between 2010 and 2013. Receiver operating characteristic curves determined the discriminatory ability of BMI, WC and WHtR to identify individuals with increased cardiometabolic risk (increased clustered triglycerides, HDL-cholesterol, systolic blood pressure, cardiorespiratory fitness, and glucose). RESULTS: A WHtR ≥ 0.5 increased the odds by 5.2 (95% confidence interval 2.6 - 10.3) of having increased cardiometabolic risk. Similar associations were observed for BMI and WC. Both BMI-z and WHtR were fair predictors of increased cardiometabolic risk, although BMI-z demonstrated the best trade-off between sensitivity and specificity, 76.1% and 63.6%, compared with 68.1% and 65.5% for WHtR. Cross-validation analysis revealed that BMI-z and WHtR correctly classified 84% of individuals (kappa score = 0.671, 95% CI 0.55, 0.79). The sensitivity of the cut-points suggests that 89.3% of individuals were correctly classified as being at risk with only 10.7% misdiagnosed whereas the specificity of the cut-points indicated that 77.8% of individuals were correctly identified as being healthy with 22.2% of individuals incorrectly diagnosed as being at risk. CONCLUSIONS: Findings suggest that WHtR provides similar cardiometabolic risk estimates to age and sex adjusted BMI.

Bid chimeras indicate that most BH3-only proteins can directly activate Bak and Bax, and show no preference for Bak versus Bax.

The mitochondrial pathway of apoptosis is initiated by Bcl-2 homology region 3 (BH3)-only members of the Bcl-2 protein family. On upregulation or activation, certain BH3-only proteins can directly bind and activate Bak and Bax to induce conformation change, oligomerization and pore formation in mitochondria. BH3-only proteins, with the exception of Bid, are intrinsically disordered and therefore, functional studies often utilize peptides based on just their BH3 domains. However, these reagents do not possess the hydrophobic membrane targeting domains found on the native BH3-only molecule. To generate each BH3-only protein as a recombinant protein that could efficiently target mitochondria, we developed recombinant Bid chimeras in which the BH3 domain was replaced with that of other BH3-only proteins (Bim, Puma, Noxa, Bad, Bmf, Bik and Hrk). The chimeras were stable following purification, and each immunoprecipitated with full-length Bcl-xL according to the specificity reported for the related BH3 peptide. When tested for activation of Bak and Bax in mitochondrial permeabilization assays, Bid chimeras were ~1000-fold more effective than the related BH3 peptides. BH3 sequences from Bid and Bim were the strongest activators, followed by Puma, Hrk, Bmf and Bik, while Bad and Noxa were not activators. Notably, chimeras and peptides showed no apparent preference for activating Bak or Bax. In addition, within the BH3 domain, the h0 position recently found to be important for Bax activation, was important also for Bak activation. Together, our data with full-length proteins indicate that most BH3-only proteins can directly activate both Bak and Bax.

Physical activity, cardiorespiratory fitness, and clustered cardiometabolic risk in 10- to 12-year-old school children: the REACH Y6 study.

OBJECTIVES: (1) Investigate whether clustered cardiometabolic risk score, cardiorespiratory fitness (CRF), sedentary time (ST), and body mass index Z-scores (BMI Z-scores), differed between participants that met and did not achieve ≥60 min of daily moderate to vigorous intensity physical activity (MVPA). (2) Compare clustered cardiometabolic risk score, BMI Z-score, ST, and MVPA by CRF status. METHODS: One hundred and one (n = 45 boys) 10- to 12-year-old participants took part in this cross-sectional study, conducted in Liverpool (Summer 2010) and Ulster (Spring 2011) UK. Assessments of blood markers, stature, sitting stature, body mass, waist circumference, flow mediated dilation (FMD), and resting blood pressure (BP) were completed. CRF (VO2 peak) was estimated using an individually calibrated treadmill protocol. Habitual MVPA and ST were assessed using an individually calibrated accelerometer protocol. Clustered cardiometabolic risk scores were calculated using blood markers, FMD (%), BP and anthropometric measures. Participants were classified as active (≥60 min MVPA) or inactive and as fit or unfit. Multivariate analysis of covariance (MANCOVA) was used to investigate differences in cardiometabolic risk, BMI Z-score, CRF, and ST by activity status. MANCOVA was also completed to assess differences in cardiometabolic risk, MVPA, ST, and BMI Z-score by fitness status. RESULTS: Inactive children exhibited significantly higher clustered cardiometabolic risk scores and ST, and lower CRF than active children. Unfit participants exhibited significantly higher clustered cardiometabolic risk scores, BMI Z-scores and ST and lower MVPA in comparison to fit participants. CONCLUSIONS: This study highlights the importance of children achieving 60 min MVPA daily and provides further evidence surrounding the importance of CRF for health.

Cardiorespiratory fitness predicts clustered cardiometabolic risk in 10-11.9-year-olds.

The aim of this study was to investigate levels of clustered cardiometabolic risk and the odds of being 'at risk' according to cardiorespiratory fitness status in children. Data from 88 10-11.9-year-old children (mean age 11.05 ± 0.51 years), who participated in either the REACH Year 6 or the Benefits of Fitness Circuits for Primary School Populations studies were combined. Waist circumference, systolic blood pressure, diastolic blood pressure, glucose, triglycerides, high-density lipoprotein cholesterol, adiponectin and C-reactive protein were assessed and used to estimate clustered cardiometabolic risk. Participants were classified as 'fit' or 'unfit' using recently published definitions (46.6 and 41.9 mL/kg/min for boys and girls, respectively), and continuous clustered risk scores between fitness groups were assessed. Participants were subsequently assigned to a 'normal' or 'high' clustered cardiometabolic risk group based on risk scores, and logistic regression analysis assessed the odds of belonging to the increased cardiometabolic risk group according to fitness. The unfit group exhibited significantly higher clustered cardiometabolic risk scores (p < 0.001) than the fit group. A clear association between fitness group and being at increased cardiometabolic risk (B = 2.509, p = 0.001) was also identified, and participants classed as being unfit were found to have odds of being classified as 'at risk' of 12.30 (95 % CI = 2.64-57.33). Conclusion Assessing cardiorespiratory fitness is a valid method of identifying children most at risk of cardiometabolic pathologies. The ROC thresholds could be used to identify populations of children most at risk and may therefore be used to effectively target a cardiometabolic risk-reducing public health intervention.

Direct visualization of Bcl-2 family protein interactions using live cell fluorescent protein redistribution assays.

Bcl-2 family proteins have important roles in tumor initiation, progression and resistance to therapy. Pro-survival Bcl-2 proteins are regulated by their interactions with pro-death BH3-only proteins making these protein-protein interactions attractive therapeutic targets. Although these interactions have been extensively characterized biochemically, there is a paucity of tools to assess these interactions in cells. Here, we address this limitation by developing quantitative, high throughput microscopy assays to characterize Bcl-2 and BH3-only protein interactions in live cells. We use fluorescent proteins to label the interacting proteins of interest, enabling visualization and quantification of their mitochondria-localized interactions. Using tool compounds, we demonstrate the suitability of our assays to characterize the cellular activity of putative therapeutic molecules that target the interaction between pro-survival Bcl-2 and pro-death BH3-only proteins. In addition to the relevance of our assays for drug discovery, we anticipate that our work will contribute to an improved understanding of the mechanisms that regulate these important protein-protein interactions within the cell.