Recombinant protein production using a Tobacco yellow dwarf virus-based episomal expression vector : control of Rep activity

Over the past decade, plants have been used as expression hosts for the production of pharmaceutically important and commercially valuable proteins. Plants offer many advantages over other expression systems such as lower production costs, rapid scale up of production, similar post-translational modification as animals and the low likelihood of contamination with animal pathogens, microbial toxins or oncogenic sequences. However, improving recombinant protein yield remains one of the greatest challenges to molecular farming. In-Plant Activation (InPAct) is a newly developed technology that offers activatable and high-level expression of heterologous proteins in plants. InPAct vectors contain the geminivirus cis elements essential for rolling circle replication (RCR) and are arranged such that the gene of interest is only expressed in the presence of the cognate viral replication-associated protein (Rep). The expression of Rep in planta may be controlled by a tissue-specific, developmentally regulated or chemically inducible promoter such that heterologous protein accumulation can be spatially and temporally controlled. One of the challenges for the successful exploitation of InPAct technology is the control of Rep expression as even very low levels of this protein can reduce transformation efficiency, cause abnormal phenotypes and premature activation of the InPAct vector in regenerated plants. Tight regulation over transgene expression is also essential if expressing cytotoxic products. Unfortunately, many tissue-specific and inducible promoters are unsuitable for controlling expression of Rep due to low basal activity in the absence of inducer or in tissues other than the target tissue. This PhD aimed to control Rep activity through the production of single chain variable fragments (scFvs) specific to the motif III of Tobacco yellow dwarf virus (TbYDV) Rep. Due to the important role played by the conserved motif III in the RCR, it was postulated that such scFvs can be used to neutralise the activity of the low amount of Rep expressed from a “leaky” inducible promoter, thus preventing activation of the TbYDV-based InPAct vector until intentional induction. Such scFvs could also offer the potential to confer partial or complete resistance to TbYDV, and possibly heterologous viruses as motif III is conserved between geminiviruses. Studies were first undertaken to determine the levels of TbYDV Rep and TbYDV replication-associated protein A (RepA) required for optimal transgene expression from a TbYDV-based InPAct vector. Transient assays in a non-regenerable Nicotiana tabacum (NT-1) cell line were undertaken using a TbYDV-based InPAct vector containing the uidA reporter gene (encoding GUS) in combination with TbYDV Rep and RepA under the control of promoters with high (CaMV 35S) or low (Banana bunchy top virus DNA-R, BT1) activity. The replication enhancer protein of Tomato leaf curl begomovirus (ToLCV), REn, was also used in some co-bombardment experiments to examine whether RepA could be substituted by a replication enhancer from another geminivirus genus. GUS expression was observed both quantitatively and qualitatively by fluorometric and histochemical assays, respectively. GUS expression from the TbYDV-based InPAct vector was found to be greater when Rep was expected to be expressed at low levels (BT1 promoter) rather than high levels (35S promoter). GUS expression was further enhanced when Rep and RepA were co-bombarded with a low ratio of Rep to RepA. Substituting TbYDV RepA with ToLCV REn also enhanced GUS expression but more importantly highest GUS expression was observed when cells were co-transformed with expression vectors directing low levels of Rep and high levels of RepA irrespective of the level of REn. In this case, GUS expression was approximately 74-fold higher than that from a non-replicating vector. The use of different terminators, namely CaMV 35S and Nos terminators, in InPAct vectors was found to influence GUS expression. In the presence of Rep, GUS expression was greater using pInPActGUS-Nos rather than pInPActGUS-35S. The only instance of GUS expression being greater from vectors containing the 35S terminator was when comparing expression from cells transformed with Rep, RepA and REnexpressing vectors and either non-replicating vectors, p35SGS-Nos or p35SGS-35S. This difference was most likely caused by an interaction of viral replication proteins with each other and the terminators. These results indicated that (i) the level of replication associated proteins is critical to high transgene expression, (ii) the choice of terminator within the InPAct vector may affect expression levels and (iii) very low levels of Rep can activate InPAct vectors hence controlling its activity is critical. Prior to generating recombinant scFvs, a recombinant TbYDV Rep was produced in E. coli to act as a control to enable the screening for Rep-specific antibodies. A bacterial expression vector was constructed to express recombinant TbYDV Rep with an Nterminal His-tag (N-His-Rep). Despite investigating several purification techniques including Ni-NTA, anion exchange, hydrophobic interaction and size exclusion chromatography, N-His-Rep could only be partially purified using a Ni-NTA column under native conditions. Although it was not certain that this recombinant N-His-Rep had the same conformation as the native TbYDV Rep and was functional, results from an electromobility shift assay (EMSA) showed that N-His-Rep was able to interact with the TbYDV LIR and was, therefore, possibly functional. Two hybridoma cell lines from mice, immunised with a synthetic peptide containing the TbYDV Rep motif III amino acid sequence, were generated by GenScript (USA). Monoclonal antibodies secreted by the two hybridoma cell lines were first screened against denatured N-His-Rep in Western analysis. After demonstrating their ability to bind N-His-Rep, two scFvs (scFv1 and scFv2) were generated using a PCR-based approach. Whereas the variable heavy chain (VH) from both cell lines could be amplified, only the variable light chain (VL) from cell line 2 was amplified. As a result, scFv1 contained VH and VL from cell line 1, whereas scFv2 contained VH from cell line 2 and VL from cell line 1. Both scFvs were first expressed in E. coli in order to evaluate their affinity to the recombinant TbYDV N-His-Rep. The preliminary results demonstrated that both scFvs were able to bind to the denatured N-His-Rep. However, EMSAs revealed that only scFv2 was able to bind to native N-His-Rep and prevent it from interacting with the TbYDV LIR. Each scFv was cloned into plant expression vectors and co-bombarded into NT-1 cells with the TbYDV-based InPAct GUS expression vector and pBT1-Rep to examine whether the scFvs could prevent Rep from mediating RCR. Although it was expected that the addition of the scFvs would result in decreased GUS expression, GUS expression was found to slightly increase. This increase was even more pronounced when the scFvs were targeted to the cell nucleus by the inclusion of the Simian virus 40 large T antigen (SV40) nuclear localisation signal (NLS). It was postulated that the scFvs were binding to a proportion of Rep, leaving a small amount available to mediate RCR. The outcomes of this project provide evidence that very high levels of recombinant protein can theoretically be expressed using InPAct vectors with judicious selection and control of viral replication proteins. However, the question of whether the scFvs generated in this project have sufficient affinity for TbYDV Rep to prevent its activity in a stably transformed plant remains unknown. It may be that other scFvs with different combinations of VH and VL may have greater affinity for TbYDV Rep. Such scFvs, when expressed at high levels in planta, might also confer resistance to TbYDV and possibly heterologous geminiviruses.

Cascading appearance-based features for visual voice activity detection

The detection of voice activity is a challenging problem, especially when the level of acoustic noise is high. Most current approaches only utilise the audio signal, making them susceptible to acoustic noise. An obvious approach to overcome this is to use the visual modality. The current state-of-the-art visual feature extraction technique is one that uses a cascade of visual features (i.e. 2D-DCT, feature mean normalisation, interstep LDA). In this paper, we investigate the effectiveness of this technique for the task of visual voice activity detection (VAD), and analyse each stage of the cascade and quantify the relative improvement in performance gained by each successive stage. The experiments were conducted on the CUAVE database and our results highlight that the dynamics of the visual modality can be used to good effect to improve visual voice activity detection performance.

Systems-based accident analysis in the led outdoor activity domain : application and evaluation of a risk management framework

Safety-compromising accidents occur regularly in the led outdoor activity domain. Formal accident analysis is an accepted means of understanding such events and improving safety. Despite this, there remains no universally accepted framework for collecting and analysing accident data in the led outdoor activity domain. This article presents an application of Rasmussen's risk management framework to the analysis of the Lyme Bay sea canoeing incident. This involved the development of an Accimap, the outputs of which were used to evaluate seven predictions made by the framework. The Accimap output was also compared to an analysis using an existing model from the led outdoor activity domain. In conclusion, the Accimap output was found to be more comprehensive and supported all seven of the risk management framework's predictions, suggesting that it shows promise as a theoretically underpinned approach for analysing, and learning from, accidents in the led outdoor activity domain.

Identifying key belief-based targets for promoting regular physical activity among mothers and fathers with young children

We investigated the key beliefs to target in interventions aimed at increasing physical activity (PA) among mothers and fathers of young children. Parents (288 mothers and 292 fathers) completed a Theory of Planned Behaviour belief-based questionnaire and a 1-week follow-up of PA behaviour. We found that a range of behavioural, normative, and control beliefs were significantly correlated with parents’ PA intentions and behaviour, with only a few differences observed in correlations between PA beliefs and intention and behaviour by gender. A range of key beliefs was identified as making independent contributions to parents’ PA intentions; however, the behavioural beliefs about improving parenting practices (β = 0.13), interfering with other commitments (β = −0.29); normative beliefs about people I exercise with (β = 0.20); and control beliefs about lack of time (β = −0.24), inconvenience (β = −0.14), lack of motivation (β = −0.34), were revealed as significant independent predictors of actual PA behaviour. Furthermore, we found that a limited amount of parents already hold these beliefs, suggesting that these key beliefs warrant changing and, therefore, are appropriate targets for subsequent intervention. The current study fills an empirical gap in the PA literature by investigating an at-risk group and using a well established theoretical framework to identify key beliefs that guide parents’ PA decision-making. Overall, we found support for parents being a unique group who hold distinctive behavioural, normative, and control beliefs toward PA. Attention to these key underlying beliefs will assist intervention work aimed at combating inactivity among this at-risk population.

MobileMums: A randomized controlled trial of an SMS-based physical activity intervention

Background Postnatal women (<12 months postpartum) are at increased risk of physical inactivity. Purpose To evaluate the efficacy and feasibility of a theory-based physical activity (PA) intervention delivered to postnatal women primarily via mobile telephone short message service (SMS). Methods Eighty-eight women were randomized to the intervention (n=45) or minimal contact control (n=43) condition. The 12-week intervention consisted of a face-to-face PA goal-setting consultation, a goal-setting magnet, three to five personally tailored SMS/week and a nominated support person who received two SMS per week. SMS content targeted constructs of social cognitive theory. Frequency (days/week) and duration (min/week) of PA participation and walking for exercise were assessed via self-report at baseline, 6 and 13 weeks. Results Intervention participants increased PA frequency by 1.82 days/week (SE±0.18) by 13 weeks (F(2,85)=4.46, p=0.038) and walking for exercise frequency by 1.08 days/ week (SE±0.24) by 13 weeks (F(2,85)=5.38, p=0.02). Positive trends were observed for duration (min/week) of PA and walking for exercise. Conclusions Intervention exposure resulted in increased frequency of PA and walking for exercise in postnatal women.

Correlates of Pedometer Use: Results from a Community-based Physical Activity Intervention Trial (10,000 Steps Rockhampton)

Background Pedometers have become common place in physical activity promotion, yet little information exists on who is using them. The multi-strategy, community-based 10,000 Steps Rockhampton physical activity intervention trial provided an opportunity to examine correlates of pedometer use at the population level. Methods Pedometer use was promoted across all intervention strategies including: local media, pedometer loan schemes through general practice, other health professionals and libraries, direct mail posted to dog owners, walking trail signage, and workplace competitions. Data on pedometer use were collected during the 2-year follow-up telephone interviews from random population samples in Rockhampton, Australia, and a matched comparison community (Mackay). Logistic regression analyses were used to determine the independent influence of interpersonal characteristics and program exposure variables on pedometer use. Results Data from 2478 participants indicated that 18.1% of Rockhampton and 5.6% of Mackay participants used a pedometer in the previous 18-months. Rockhampton pedometer users (n = 222) were more likely to be female (OR = 1.59, 95% CI: 1.11, 2.23), aged 45 or older (OR = 1.69, 95% CI: 1.16, 2.46) and to have higher levels of education (university degree OR = 4.23, 95% CI: 1.86, 9.6). Respondents with a BMI > 30 were more likely to report using a pedometer (OR = 1.68, 95% CI: 1.11, 2.54) than those in the healthy weight range. Compared with those in full-time paid work, respondents in 'home duties' were significantly less likely to report pedometer use (OR = 0.18, 95% CI: 0.06, 0.53). Exposure to individual program components, in particular seeing 10,000 Steps street signage and walking trails or visiting the website, was also significantly associated with greater pedometer use. Conclusion Pedometer use varies between population subgroups, and alternate strategies need to be investigated to engage men, people with lower levels of education and those in full-time 'home duties', when using pedometers in community-based physical activity promotion initiatives.

A rebirth of physical activity in parents : applying a theory-based approach

Physical inactivity is a serious concern both nationally and internationally. Despite the numerous benefits of performing regular physical activity, many individuals lead sedentary lifestyles. Of concern, though, is research showing that some population sub-groups are less likely to be active, such as parents of young children. Although there is a vast amount of research dedicated to understanding people.s physical activity-related behaviours, there is a paucity of research examining those factors that influence parental physical activity. More importantly, research applying theoretical models to understand physical activity decision-making among this at-risk population is limited. Given the current obesity epidemic, the decline in physical activity with parenthood, and the many social and health benefits associated with regular physical activity, it is important that adults with young children are sufficiently active. In light of the dearth of research examining parental physical activity and the scant research applying a theory-based approach to gain this understanding, the overarching aim of the current program of research was to adopt a mixed methods approach as well as use sound theoretical frameworks to understand the regular physical activity behaviour of mothers and fathers with young children. This program of research comprised of three distinct stages: a qualitative stage exploring individual, social, and psychological factors that influence parental regular physical activity (Stage 1); a quantitative stage identifying the important predictors of parental regular physical activity intentions and behaviour using sound theoretical frameworks and testing a single-item measure for assessing parental physical activity behaviour (Stage 2); and a qualitative stage exploring strategies for an intervention program aimed at increasing parental regular physical activity (Stage 3). As a thesis by publication, eight papers report the findings of this program of research; these papers are presented according to the distinct stages of investigation that guided this program of research. Stage One of the research program comprised a qualitative investigation using a focus group/interview methodology with parents of children younger than 5 years of age (N = 40; n = 21 mothers, n = 19 fathers) (Papers 1, 2, and 3). Drawing broadly on a social constructionist approach (Paper 1), thematic analytic methods revealed parents. understandings of physical activity (e.g., requires effort), patterns of physical activity-related behaviours (e.g., grab it when you can, declining physical activity habits), and how constructions of social role expectations might influence parents. physical activity decision making (e.g., creating an active family culture, guilt and selfishness). Drawing on the belief-based framework of the TPB (Paper 2), thematic content analytic methods revealed parents. commonly held beliefs about the advantages (e.g., improves parenting practices), disadvantages (e.g., interferes with commitments), barriers (e.g., time), and facilitators (e.g., social support) to performing regular physical activity. Parents. normative beliefs about social approval from important others or groups (e.g., spouse/partner) were also identified. Guided by theories of social support, Paper Three identified parents. perceptions about the specific social support dimensions that influence their physical activity decision making. Thematic content analysis identified instrumental (e.g., providing childcare, taking over chores), emotional (e.g., encouragement, companionship), and informational support (e.g., ideas and advice) as being important to the decision-making of parents in relation to their regular physical activity behaviour. The results revealed also that having support for being active is not straightforward (e.g., guilt-related issues inhibited the facilitative nature of social support for physical activity). Stage Two of the research program comprised a quantitative examination of parents. physical activity intentions and behaviour (Papers 4, 5, 6, and 7). Parents completed an extended TPB questionnaire at Time 1 (N = 580; n = 288 mothers, n = 292 fathers) and self-reported their physical activity at Time 2, 1 week later (N = 458; n = 252 mothers, n = 206 fathers). Paper Four revealed key behavioural (e.g., improving parenting practices), normative (e.g., people I exercise with), and control (e.g., lack of time) beliefs as significant independent predictors of parental physical activity. A test of the TPB augmented to include the constructs of self-determined motivation and planning was assessed in Paper Five. The findings revealed that the effect of self-determined motivation on intention was fully mediated by the TPB variables and the impact of intention on behaviour was partially mediated by the planning variables. Slight differences in the model.s motivational sequence between the sexes were also noted. Paper Six investigated, within a TPB framework, a range of social influences on parents. intentions to be active. For both sexes, attitude, perceived behavioural control, group norms, friend general support, and an active parent identity predicted intentions, with subjective norms and family support further predicting mothers. intentions and descriptive norms further predicting fathers. intentions. Finally, the measurement of parental physical activity was investigated in Paper Seven of Stage Two. The results showed that parents are at risk of low levels of physical activity, with the findings also revealing validation support for a brief single-item physical activity measure. Stage Three of the research program comprised a qualitative examination of parents. (N = 12; n = 6 mothers, n = 6 fathers) ideas for strategies that may be useful for developing and delivering an intervention program aimed at increasing parental physical activity (Paper 8). Parents revealed a range of strategies for what to include in a physical activity intervention designed for parents of young children. For example, parents identified persuasion and information type messages, problem-solving strategies that engage parents in generating a priority list of their lifestyle commitments, and behavioural modification techniques such as goal setting and incentives. Social intervention strategies (e.g., social comparison, counselling) and environmental approaches (e.g., community-based integrative parent/child programs) were also identified as was a skill-based strategy in helping parents generate a flexible life/family plan. Additionally, a range of strategies for how to best deliver a parental physical activity intervention was discussed. Taken as a whole, Paper Eight found that adopting a multifaceted approach in both the design and implementation of a resultant physical activity intervention may be useful in helping to increase parental physical activity. Overall, this program of research found support for parents as a unique group who hold both similar and distinctive perceptions about regular physical activity to the general adult population. Thus, these findings highlight the importance of targeting intervention strategies for parents of young children. Additionally, the findings suggest that it might also be useful to tailor some messages specifically to each sex. Effective promotion of physical activity in parents of young children is essential given the low rate of activity in this population. Results from this program of research highlight parents as an at-risk group for inactivity and provide an important first step in identifying the factors that influence both mothers. and fathers. physical activity decision making. These findings, in turn, provide a foundation on which to build effective intervention programs aimed at increasing parents. regular physical activity which is essential for ensuring the health and well-being of parents with young children.

Incentive-based interventions for increasing physical activity and fitness

This is the protocol for a review and there is no abstract. The objectives are as follows: The main aim of the review is to determine the effectiveness of using incentive-based approaches (IBAs) (financial and non-financial) to increase physical activity in community-dwelling children and adults. A secondary objective will be to address the use of incentives to improve cardiovascular and metabolic fitness. A final objective will be to explore: - whether there are any adverse effects associated with the use of IBAs for increasing physical activity; - whether there are any differential effects of IBAs within and between study populations by age, gender, education, inequalities and health status; and - whether the use of disincentive/aversive approaches leads to a reduction in sedentary behaviour.

Australian adolescents’ perceived school-based barriers and facilitators to engagement in physical activity

OBJECTIVE: School-aged youth spend a significant amount of time either in transit to and from school, or within school settings performing a range of varying learning-based activities. Adolescent physical activity has also been shown to increase the likelihood of maintaining physical activity throughout adulthood. The purpose of this study is to investigate adolescents’ perceived school-based barriers and facilitators to engagement in physical activity. METHODS: One-hundred and twenty four participants (38 males and 86 females) were recruited from two non-denominational same-sex private schools, in Brisbane, Australia. The mean age and standard deviation (SD) was 13.83 (0.56) and 14.40 (2.33) for males and females respectively. Participants responded to a series questions regarding perceived barriers and facilitators to engagement in physical activity. Quantitative data was analysed using descriptive statistics and frequency distributions, and qualitative data with thematic analysis. RESULTS: A total of 121 (97.6%) participants had complete data sets and were included in the analysis. School timetable (44.6%), homework (81.8%), and assessment (81.0%) were identified as the most prominent perceived factors, increasing the difficulty of physical activity engagement. Physical Education classes (71.9%) and school sport programs (80.2%) were identified as the most prominent perceived factors that facilitate engagement in physical activity. There was no significant gender effect. CONCLUSIONS: Each of the identified factors perceived by adolescent's as either barriers or facilitators to engagement in physical activity may be addressed by administrators at a school and government policy level. These may include strategies such as; increasing the assigned hours to physical education classes, providing additional extra-curricular sporting opportunities, and reviewing the time allocated to homework and assessment items. This may provide a simpler, low-cost solution to increasing youth physical activity, as opposed to contemporary higher-cost strategies utilising increased staff commitment, mass media, provision of equipment and counsellors and other health professionals.

Pre-service Physical Education teachers and concerns regarding their ability to successfully facilitate school-based physical activity promotion strategies

Regular physical activity (PA) in youth has numerous immediate and long-term health benefits. With several studies indicating low levels of youth PA globally, schools settings have become increasingly critical settings for youth health promotion strategies. The role of physical education (PE) teachers has long been considered central to the facilitation of such strategies. However, PE teachers have a selfreported lack of knowledge, skills, understanding, and competence to successfully implement these strategies. Tertiary education programs are fundamental to adequately preparing, and shaping the attitudes and philosophies of future PE teachers towards their involvement within these programs. The aim of this investigation was to explore the beliefs and perceptions of future secondary school PE teachers, regarding their potential roles in future school-based programs designed to promote student PA. Fifty-seven (21 males and 36 females) pre-service PE teachers completed a series of open-ended survey questions concerning their perceptions towards participating in school-based PA promotion programs both as preservice during practicum, and prospectively as practising teachers. Responses were analysed thematically. Participants responded both positively and enthusiastically to both questions. Concerns regarding time, and the intention or expectation to participate in such programs were also key themes for pre-service and practicing teacher participation respectively. Critically in this study, participants did not identify any limitations which may impact upon their ability to successfully promote youth PA in school settings. This may indicate that participants have misconceptions regarding their ability to fulfil this role, or conversely, the deficiency of current PE teachers regarding school-based PA promotion has been recognised by the tertiary institution, and addressed to adequately prepare its students. School-based PA promotion is an integral element of pre-service PE teacher education, and ongoing professional development of practicing PE teachers. This trend is expected to continue in the future, in order to address ongoing public health concerns.

Home-based activity program for older people with depressive symptoms: DeLLITE–A randomized controlled trial

PURPOSE We wanted to assess the effectiveness of a home-based physical activity program, the Depression in Late Life Intervention Trial of Exercise (DeLLITE), in improving function, quality of life, and mood in older people with depressive symptoms. METHODS We undertook a randomized controlled trial involving 193 people aged 75 years and older with depressive symptoms at enrollment who were recruited from primary health care practices in Auckland, New Zealand. Participants received either an individualized physical activity program or social visits to control for the contact time of the activity intervention delivered over 6 months. Primary outcome measures were function, a short physical performance battery comprising balance and mobility, and the Nottingham Extended Activities of Daily Living scale. Secondary outcome measures were quality of life, the Medical Outcomes Study 36-item short form, mood, Geriatric Depression Scale (GDS-15), physical activity, Auckland Heart Study Physical Activity Questionnaire, and self-report of falls. Repeated measures analyses tested the differential impact on outcomes over 12 months’ follow-up. RESULTS The mean age of the participants was 81 years, and 59% were women. All participants scored in the at–risk category on the depression screen, 53% had a Diagnostic and Statistical Manual of Mental Disorders or International Classification of Diseases, Tenth Revision diagnosis of major depression or scored more than 4 on the GDS-15 at baseline, indicating moderate or severe depression. Almost all participants, 187 (97%), completed the trial. Overall there were no differences in the impact of the 2 interventions on outcomes. Mood and mental health related quality of life improved for both groups. CONCLUSION he DeLLITE activity program improved mood and quality of life for older people with depressive symptoms as much as the effect of social visits. Future social and activity interventions should be tested against a true usual care control.

Semi-binary based video features for activity representation

Efficient and effective feature detection and representation is an important consideration when processing videos, and a large number of applications such as motion analysis, 3D scene understanding, tracking etc. depend on this. Amongst several feature description methods, local features are becoming increasingly popular for representing videos because of their simplicity and efficiency. While they achieve state-of-the-art performance with low computational complexity, their performance is still too limited for real world applications. Furthermore, rapid increases in the uptake of mobile devices has increased the demand for algorithms that can run with reduced memory and computational requirements. In this paper we propose a semi binary based feature detectordescriptor based on the BRISK detector, which can detect and represent videos with significantly reduced computational requirements, while achieving comparable performance to the state of the art spatio-temporal feature descriptors. First, the BRISK feature detector is applied on a frame by frame basis to detect interest points, then the detected key points are compared against consecutive frames for significant motion. Key points with significant motion are encoded with the BRISK descriptor in the spatial domain and Motion Boundary Histogram in the temporal domain. This descriptor is not only lightweight but also has lower memory requirements because of the binary nature of the BRISK descriptor, allowing the possibility of applications using hand held devices.We evaluate the combination of detectordescriptor performance in the context of action classification with a standard, popular bag-of-features with SVM framework. Experiments are carried out on two popular datasets with varying complexity and we demonstrate comparable performance with other descriptors with reduced computational complexity.

Conducting accelerometer-based activity assessments in field-based research

Purpose The purpose of this review is to address important methodological issues related to conducting accelerometer-based assessments of physical activity in free-living individuals. Methods We review the extant scientific literature for empirical information related to the following issues: product selection, number of accelerometers needed, placement of accelerometers, epoch length, and days of monitoring required to estimate habitual physical activity. We also discuss the various options related to distributing and collecting monitors and strategies to enhance compliance with the monitoring protocol. Results No definitive evidence exists currently to indicate that one make and model of accelerometer is more valid and reliable than another. Selection of accelerometer therefore remains primarily an issue of practicality, technical support, and comparability with other studies. Studies employing multiple accelerometers to estimate energy expenditure report only marginal improvements in explanatory power. Accelerometers are best placed on hip or the lower back. Although the issue of epoch length has not been studied in adults, the use of count cut points based on 1-min time intervals maybe inappropriate in children and may result in underestimation of physical activity. Among adults, 3–5 d of monitoring is required to reliably estimate habitual physical activity. Among children and adolescents, the number of monitoring days required ranges from 4 to 9 d, making it difficult to draw a definitive conclusion for this population. Face-to-face distribution and collection of accelerometers is probably the best option in field-based research, but delivery and return by express carrier or registered mail is a viable option. Conclusion Accelerometer-based activity assessments requires careful planning and the use of appropriate strategies to increase compliance.