A method for conducting suppression assays using small numbers of tissue-isolated regulatory T cells

The suppression assay is a commonly performed assay, measuring the ability of regulatory T cells (Treg) to suppress T cell proliferation. Most frequently, Treg are obtained from the peripheral blood or spleen. Lower yields are obtained by isolation from other tissues, rendering downstream suppression assays challenging to perform. Furthermore, the importance of suppressive subpopulations of Treg favours their isolation by fluorescent-activated cell sorting. Here we describe a method to isolate Treg from human tissues, using colorectal cancer tissue as an example. Treg suppressive capacity was further examined by expression of CCR5 to demonstrate the ability of our method to assess the suppressive capacity of regulatory T cell subsets. To optimise the standard suppression assay to achieve our research aims, the following modifications were made: • Treg, isolated from tissues, were sorted directly into a well-plate. • Responder T cells, which had been fluorescently-labelled prior to sorting, were added directly into the well-plate. • Human Treg Suppression Inspector beads (Miltenyi Biotec Ltd., UK) provided a polyclonal stimulus for proliferation and were added to each well at a bead:lymphocyte ratio of 1:2. This method quantified the suppression of responder T cell proliferation by small numbers of strictly-defined Treg populations isolated from tissues

Gotta catch 'em all or not enough time: Users motivations for playing Pokemon Go (TM) and non-users' reasons for not installing

Urban exergames are played in the real-world environment using built-in mobile phone sensors. The influence of Pokémon Go on physical activity and sitting time has been examined previously, however limited research has explored motivations for using the application. Thus, the aim of this study was to explore Pokémon Go users' motivations for using the application, exusers' reasons for abandoning the game and non-users' reasons for not installing. After institutional ethical approval, the 'Physical Activity and Pokémon Go' questionnaire was developed using QualtricsTM and distributed using social media soon after launch in the United Kingdom (baseline). At baseline a total of 461 participants (n=193 male, n=265 female, n=3 transgender) who were predominantly white (n=420) and did not self-report a disability (n=443) completed the questionnaire. Users' (n=236) were questioned on their motivations for using Pokémon Go and non-users' provided reasons for not installing. At 3 months a total of 127 participants (n=23 users) completed the questionnaire again and all qualitative data was thematically analyzed. The most commonly reported reason for using Pokémon Go was 'to have fun' which was 86% and 83% at baseline and 3 months respectively. The second most frequent reason at baseline was 'friends were using it' (58%) and at 3 months was 'to be outside' (48%). The least common motivation for using Pokémon Go at both baseline and 3 months was 'to meet new people' (8% and 0% respectively). At baseline, social motives and competition were two general themes which encapsulated Pokémon Go users' motivation for using the application. There were two general themes reported by Pokémon Go users' as to why they did not think they would use the application in the future. These were application related factors and factors unrelated to Pokémon Go. Non-users reported a range of reasons for not using Pokémon Go, including lack of interest and a lack of time. Safety concerns and risk of adverse events were reported by both users and non-users. This is the first study to thematically analyze motives for using Pokémon Go in which the findings are: 1) future smartphone applications aiming to increase physical activity must ensure that objectives evolve to maintain initial interest and motivation to engage with applications; 2) game developers must consider the required phone storage and capability which could be a barrier to downloading; and 3) concerns of using the application including the safety of users and those around them

Examining consumers’ adoption of wearable healthcare technology: The role of health attributes

With the advancement of information technology, wearable healthcare technology has emerged as one of the promising technologies to improve the wellbeing of individuals. However, the adoption of wearable healthcare technology has lagged when compared to other well-established durable technology products, such as smartphones and tablets, because of the inadequate knowledge of the antecedents of adoption intention. The aim of this paper is to address an identified gap in the literature by empirically testing a theoretical model for examining the impact of consumers’ health beliefs, health information accuracy, and the privacy protection of wearable healthcare technology on perceived usefulness. Importantly, this study also examines the influences of perceived usefulness, consumer innovativeness, and reference group influence on the adoption intention of wearable healthcare technology. The model seeks to enhance understanding of the influential factors in adopting wearable healthcare technology. Finally, suggestions for future research for the empirical investigation of the model are provided

Development and evaluation of a diagnostic cytokine-release assay for Mycobacterium suricattae infection in meerkats (Suricata suricatta)

CITATION: Clarke, C., et al. 2017. Development and evaluation of a diagnostic cytokine-release assay for mycobacterium suricattae infection in meerkats (Suricata suricatta). BMC Veterinary Research, 13:2, doi:10.1186/s12917-016-0927-x.The original publication is available at http://bmcvetres.biomedcentral.comBackground: Sensitive diagnostic tools are necessary for the detection of Mycobacterium suricattae infection in meerkats (Suricata suricatta) in order to more clearly understand the epidemiology of tuberculosis and the ecological consequences of the disease in this species. We therefore aimed to develop a cytokine release assay to measure antigen-specific cell-mediated immune responses of meerkats. Results: Enzyme-linked immunosorbent assays (ELISAs) were evaluated for the detection of interferon-gamma (IFN-γ) and IFN-γ inducible protein 10 (IP-10) in meerkat plasma. An IP-10 ELISA was selected to measure the release of this cytokine in whole blood in response to Bovigam® PC-HP Stimulating Antigen, a commercial peptide pool of M. bovis antigens. Using this protocol, captive meerkats with no known M. suricattae exposure (n = 10) were tested and results were used to define a diagnostic cut off value (mean plus 2 standard deviations). This IP-10 release assay (IPRA) was then evaluated in free-living meerkats with known M. suricattae exposure, categorized as having either a low, moderate or high risk of infection with this pathogen. In each category, respectively, 24.7%, 27.3% and 82.4% of animals tested IPRA-positive. The odds of an animal testing positive was 14.0 times greater for animals with a high risk of M. suricattae infection compared to animals with a low risk. Conclusion: These results support the use of this assay as a measure of M. suricattae exposure in meerkat populations. Ongoing longitudinal studies aim to evaluate the value of the IPRA as a diagnostic test of M. suricattae infection in individual animals.http://bmcvetres.biomedcentral.com/articles/10.1186/s12917-016-0927-xPublisher's versio

Effects of physical activity on functional health of older adults: a systematic review

Reviews on the relationships between functional health and physical activity of general older adults have been well documented in literature. However, specific age range of older adults, in particular, older adults of 75 years or above, is currently under-examined. A systematic review was conducted to investigate the effects of physical activity on functional health older adults aged 75 years or above. The reviewed articles cover a variety range of functional health outcomes, including balance, muscle conditioning, joint range of motion, quadriceps strength, reaction time, gait speed, health-related quality of life, back and knee pain, muscle mass, and walking ability. In general, interventions of the reviewed articles had favourable effects on function health of older adults. While physical activity has been identified as an important determinant of functional health, the ways to engage in and accumulate sufficient daily physical activity warrant investigation. It is also important to explore interventions which enhance daily, self-driven physical activity of elderly, as normally supervised physical activity bears higher costs

Proper Distance Metrics for Phylogenetic Analysis Using Complete Genomes without Sequence Alignment

A shortcoming of most correlation distance methods based on the composition vectors without alignment developed for phylogenetic analysis using complete genomes is that the “distances” are not proper distance metrics in the strict mathematical sense. In this paper we propose two new correlation-related distance metrics to replace the old one in our dynamical language approach. Four genome datasets are employed to evaluate the effects of this replacement from a biological point of view. We find that the two proper distance metrics yield trees with the same or similar topologies as/to those using the old “distance” and agree with the tree of life based on 16S rRNA in a majority of the basic branches. Hence the two proper correlation-related distance metrics proposed here improve our dynamical language approach for phylogenetic analysis

Ipl1/aurora kinase suppresses S-CDK-driven spindle formation during prophase I to ensure chromosome integrity during meiosis

Cells coordinate spindle formation with DNA repair and morphological modifications to chromosomes prior to their segregation to prevent cell division with damaged chromosomes. Here we uncover a novel and unexpected role for Aurora kinase in preventing the formation of spindles by Clb5-CDK (S-CDK) during meiotic prophase I and when the DDR is active in budding yeast. This is critical since S-CDK is essential for replication during premeiotic S-phase as well as double-strand break induction that facilitates meiotic recombination and, ultimately, chromosome segregation. Furthermore, we find that depletion of Cdc5 polo kinase activity delays spindle formation in DDR-arrested cells and that ectopic expression of Cdc5 in prophase I enhances spindle formation, when Ipl1 is depleted. Our findings establish a new paradigm for Aurora kinase function in both negative and positive regulation of spindle dynamics

Balancing the dilution and oddity effects: Decisions depend on body size

Background Grouping behaviour, common across the animal kingdom, is known to reduce an individual's risk of predation; particularly through dilution of individual risk and predator confusion (predator inability to single out an individual for attack). Theory predicts greater risk of predation to individuals more conspicuous to predators by difference in appearance from the group (the ‘oddity’ effect). Thus, animals should choose group mates close in appearance to themselves (eg. similar size), whilst also choosing a large group. Methodology and Principal Findings We used the Trinidadian guppy (Poecilia reticulata), a well known model species of group-living freshwater fish, in a series of binary choice trials investigating the outcome of conflict between preferences for large and phenotypically matched groups along a predation risk gradient. We found body-size dependent differences in the resultant social decisions. Large fish preferred shoaling with size-matched individuals, while small fish demonstrated no preference. There was a trend towards reduced preferences for the matched shoal under increased predation risk. Small fish were more active than large fish, moving between shoals more frequently. Activity levels increased as predation risk decreased. We found no effect of unmatched shoal size on preferences or activity. Conclusions and Significance Our results suggest that predation risk and individual body size act together to influence shoaling decisions. Oddity was more important for large than small fish, reducing in importance at higher predation risks. Dilution was potentially of limited importance at these shoal sizes. Activity levels may relate to how much sampling of each shoal was needed by the test fish during decision making. Predation pressure may select for better decision makers to survive to larger size, or that older, larger fish have learned to make shoaling decisions more efficiently, and this, combined with their size relative to shoal-mates, and attractiveness as prey items influences shoaling decisions