Understanding nanoparticle uptake by biological cells is fundamentally important to wide-ranging fields from nanotoxicology to drug delivery. It is now accepted that the arrival of nanoparticles at the cell is an extremely complicated process, shaped by many factors including unique nanoparticle physico-chemical characteristics, protein-particle interactions and subsequent agglomeration, diffusion and sedimentation. Sequentially, the nanoparticle internalisation process itself is also complex, and controlled by multiple aspects of a cell’s state. Despite this multitude of factors, here we demonstrate that the statistical distribution of the nanoparticle dose per endosome is independent of the initial administered dose and exposure duration. Rather, it is the number of nanoparticle containing endosomes that are dependent on these initial dosing conditions. These observations explain the heterogeneity of nanoparticle delivery at the cellular level and allow the derivation of simple, yet powerful probabilistic distributions that accurately predict the nanoparticle dose delivered to individual cells across a population.
Physical inactivity is one of the most prevalent risk factors for non-communicable diseases in the world. A fundamental barrier to enhancing physical activity levels and decreasing sedentary behavior is limited by our understanding of associated measurement and analytical techniques. The number of analytical techniques for physical activity measurement has grown significantly, and although emerging techniques may advance analyses, little consensus is presently available and further synthesis is therefore required. The objective of this review was to identify the accuracy of emerging analytical techniques used for physical activity measurement in humans. We conducted a search of electronic databases using Web of Science, PubMed, and Google Scholar. This review included studies written in English and published between January 2010 and December 2014 that assessed physical activity using emerging analytical techniques and reported technique accuracy. A total of 2064 papers were initially retrieved from three databases. After duplicates were removed and remaining articles screened, 50 full-text articles were reviewed, resulting in the inclusion of 11 articles that met the eligibility criteria. Despite the diverse nature and the range in accuracy associated with some of the analytic techniques, the rapid development of analytics has demonstrated that more sensitive information about physical activity may be attained. However, further refinement of these techniques is needed.
Obese children move less and with greater difficulty than their normal-weight counterparts. Whilst the effect of high BMI on cardiovascular fitness is well known, the effect on movement quality characteristics during a standardised fitness test has not been investigated. The aims of this study were, to characterise the movement quality of children performing the multi-stage fitness test (MSFT), and, report how movement quality characteristics cluster according to weight status. One hundred and three children (10.3±0.6 y, 1.42±0.08m, 37.8±9.3kg, BMI; 18.5±3.3kgm(2)) performed the MSFT whilst wearing an ankle mounted accelerometer. BMI groups were used to classify children as underweight (UW), normal weight (NW), overweight (OW) and obese (OB). Characteristics of movement were profiled using a clustering algorithm. Spearman's rho was used to assess relationship with BMI group, and a Mann-Whitney U test was used to assess differences between BMI groups. Obese children had significantly lower spectral purity than every other group and significantly lower time to exhaustion (TTE) than UW and NW children (P<0.05). BMI was clustered with stride profile and TTE with spectral purity. Significant negative correlations (P<0.05) were found between BMI and TTE (r=-0.25), spectral purity (r=-0.24), integrated acceleration (r=-0.22), stride angle (r=-0.23) and stride variability (r=-0.22). This was the first study to report the spectral purity of children's gait. Further analysis unveiled key performance characteristics that differed between BMI groups. These were (i) representative of children's performance during the MSFT and, (ii) significantly negatively correlated with BMI.
By using multi-dimensional analysis of similarity measures between participants rather than direct parameterisation of the physiological data, complex and varied patterns of physical motion can be quantified, allowing objective and robust profiling of relative function across participant groups.
Immunofluorescence microscopy is an essential tool for tissue‐based research, yet data reporting is almost always qualitative. Quantification of images, at the per‐cell level, enables “flow cytometry‐type” analyses with intact locational data but achieving this is complex. Gastrointestinal tissue, for example, is highly diverse: from mixed‐cell epithelial layers through to discrete lymphoid patches. Moreover, different species (e.g., rat, mouse, and humans) and tissue preparations (paraffin/frozen) are all commonly studied. Here, using field‐relevant examples, we develop open, user‐friendly methodology that can encompass these variables to provide quantitative tissue microscopy for the field. Antibody‐independent cell labeling approaches, compatible across preparation types and species, were optimized. Per‐cell data were extracted from routine confocal micrographs, with semantic machine learning employed to tackle densely packed lymphoid tissues. Data analysis was achieved by flow cytometry‐type analyses alongside visualization and statistical definition of cell locations, interactions and established microenvironments. First, quantification of Escherichia coli passage into human small bowel tissue, following Ussing chamber incubations exemplified objective quantification of rare events in the context of lumen‐tissue crosstalk. Second, in rat jejenum, precise histological context revealed distinct populations of intraepithelial lymphocytes between and directly below enterocytes enabling quantification in context of total epithelial cell numbers. Finally, mouse mononuclear phagocyte—T cell interactions, cell expression and significant spatial cell congregations were mapped to shed light on cell–cell communication in lymphoid Peyer's patch. Accessible, quantitative tissue microscopy provides a new window‐of‐insight to diverse questions in gastroenterology. It can also help combat some of the data reproducibility crisis associated with antibody technologies and over‐reliance on qualitative microscopy. © 2020 The Authors. Cytometry Part A published by Wiley Periodicals LLC. on behalf of International Society for Advancement of Cytometry.
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