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1.
Memory ; : 1-14, 2024 Jul 04.
Article in English | MEDLINE | ID: mdl-38963905

ABSTRACT

According to Conway's view, Autobiographical memory (AM) construction is accompanied by control processes. These processes range from filtering out relevant memories according to the current context, to generating or elaborating appropriate retrieval cues. These processes can be conceptualised as metacognition, the ability to control and monitor cognitive processes. Experimentally, little has been carried out to support the idea that metacognition is involved in AM. To assess this, we designed a task, the Feeling of Retrieval. Participants had to predict whether cue words would facilitate AM access (i.e., fluent access cues) or not (i.e., limited access cues) in a limited time (either 1 (Exp. 2) or 2 (Exp. 1) s). Later, they retrieved memories in response to both types of cues. Results show that cues judged as fluent access led to better AM generation, as illustrated by AM retrieval latency and a subjective measure of the ease with which the AMs were retrieved. These rapid predictions may rely on epistemic feelings and / or other mnemonic cues such as a partial retrieval of information. This metacognitive access to the earliest stages of AM retrieval illustrates the ability to monitor AM processes as proposed by Conway (2005).

2.
New Phytol ; 240(3): 1305-1326, 2023 11.
Article in English | MEDLINE | ID: mdl-37678361

ABSTRACT

Pollen and tracheophyte spores are ubiquitous environmental indicators at local and global scales. Palynology is typically performed manually by microscopic analysis; a specialised and time-consuming task limited in taxonomical precision and sampling frequency, therefore restricting data quality used to inform climate change and pollen forecasting models. We build on the growing work using AI (artificial intelligence) for automated pollen classification to design a flexible network that can deal with the uncertainty of broad-scale environmental applications. We combined imaging flow cytometry with Guided Deep Learning to identify and accurately categorise pollen in environmental samples; here, pollen grains captured within c. 5500 Cal yr BP old lake sediments. Our network discriminates not only pollen included in training libraries to the species level but, depending on the sample, can classify previously unseen pollen to the likely phylogenetic order, family and even genus. Our approach offers valuable insights into the development of a widely transferable, rapid and accurate exploratory tool for pollen classification in 'real-world' environmental samples with improved accuracy over pure deep learning techniques. This work has the potential to revolutionise many aspects of palynology, allowing a more detailed spatial and temporal understanding of pollen in the environment with improved taxonomical resolution.


Subject(s)
Deep Learning , Artificial Intelligence , Flow Cytometry , Phylogeny , Pollen
3.
J Geriatr Oncol ; 14(3): 101449, 2023 04.
Article in English | MEDLINE | ID: mdl-36870223

ABSTRACT

INTRODUCTION: The COVID-19 pandemic impacted the care and experiences of people with cancer, but it presented an opportunity to improve the delivery of outpatient care post-pandemic. MATERIALS AND METHODS: We performed an observational cross-sectional study with people with lung cancer throughout the COVID-19 pandemic. A survey investigated patients' experiences and preferences regarding the delivery of cancer care to plan for post-pandemic care, as well as the pandemic's impact on their functional status (physical and psycho-social), exploring the role of age and frailty. RESULTS: Amongst 282 eligible participants, 88%, 86%, and 59% of patients reported feeling appropriately supported during the pandemic by their cancer centre, friends/family, and primary care services, respectively. Remote oncology consultations were delivered to 90% of patients during the pandemic, of which 3% did not meet patients' expectations. Regarding post-pandemic outpatient care preferences, face-to-face appointments were preferred by 93% for the first appointment, by 64% when discussing imaging results, and by 60% for reviews during anti-cancer treatments. Older patients aged 70 years and above were more likely to favour face-to-face appointments (p = 0.007), regardless of their frailty status. Patient preferences changed over time, with the more recent participants preferring remote appointments during anti-cancer treatments (p = 0.0278). Regarding the pandemic's impact, abnormal levels of anxiety and depression were found in 16% and 17% of patients, respectively. Younger patients experienced higher abnormal levels of anxiety and depression (p = 0.036, p = 0.021). Amongst the older sub-group, those with frailty had higher levels of anxiety and depression (p < 0.001). Amongst all participants, 54% reported a considerable negative impact from the pandemic on different aspects of their daily life, particularly emotional and psychological health and sleep patterns, which were more marked in younger patients and the older sub-group with frailty. Older patients without frailty reported the least impact on their functional status. DISCUSSION: There is a need for more personalised outpatient consultation options during cancer care. Whilst there is a preference for face-to-face consultations for older patients, following the pandemic there is a growing acceptance of remote consultations particularly during anti-cancer treatment. Older patients with lung cancer without frailty were less affected by the pandemic than those with frailty and younger patients, requiring less support from healthcare services.


Subject(s)
COVID-19 , Frailty , Lung Neoplasms , Humans , Cross-Sectional Studies , Pandemics , Outpatients , Frailty/epidemiology , Lung Neoplasms/epidemiology , Lung Neoplasms/therapy , Ambulatory Care
4.
Cell Rep Methods ; 3(2): 100398, 2023 02 27.
Article in English | MEDLINE | ID: mdl-36936072

ABSTRACT

Unlocking and quantifying fundamental biological processes through tissue microscopy requires accurate, in situ segmentation of all cells imaged. Currently, achieving this is complex and requires exogenous fluorescent labels that occupy significant spectral bandwidth, increasing the duration and complexity of imaging experiments while limiting the number of channels remaining to address the study's objectives. We demonstrate that the excitation light reflected during routine confocal microscopy contains sufficient information to achieve accurate, label-free cell segmentation in 2D and 3D. This is achieved using a simple convolutional neural network trained to predict the probability that reflected light pixels belong to either nucleus, cytoskeleton, or background classifications. We demonstrate the approach across diverse lymphoid tissues and provide video tutorials demonstrating deployment in Python and MATLAB or via standalone software for Windows.


Subject(s)
Image Processing, Computer-Assisted , Imaging, Three-Dimensional , Image Processing, Computer-Assisted/methods , Imaging, Three-Dimensional/methods , Microscopy, Confocal/methods , Neural Networks, Computer , Software
5.
J Pediatr Gastroenterol Nutr ; 75(2): e20-e24, 2022 08 01.
Article in English | MEDLINE | ID: mdl-35666860

ABSTRACT

OBJECTIVE: The incidence of paediatric inflammatory bowel disease (IBD) has been increasing over 25 years; however, contemporary trends are not established and the impact of COVID-19 on case rates is unclear. METHODS: Data from Southampton Children's hospital prospective IBD database were retrieved for 2002-2021. Incidence rates were calculated based on referral area populations and temporal trends analysed. Disease prevalence for those aged <18 years was calculated for 2017-2021. Monoclonal prescriptions were reported. RESULTS: In total, 1150 patients were included (mean age at diagnosis 12.63 years, 40.5% female). An estimated 704 patients had Crohn's disease (61.2%), 385 had ulcerative colitis (33.5%), and 61 had IBD unclassified (5.3%). Overall IBD incidence increased, ß = 0.843, P = 3 × 10 -6 , driven by Crohn's disease, ß = 0.732, P = 0.00024 and ulcerative colitis, ß = 0.816, P = 0.000011. There was no change in IBDU incidence, ß = 0.230, P = 0.33. From 2002-2021, 51 patients were diagnosed <6 years of age, 160 patients aged 6 to <10 years and 939 patients aged 10 to <18 years of age. Increased incidence was observed in patients aged 10 to <18 years of age (ß = 0.888, P = 1.8 × 10 -7 ). There was no significant change in incidence of IBD in <6 years (ß = 0.124, P = 0.57), or 6 to <10 years (ß = 0.146, P = 0.54). IBD prevalence increased by an average of 1.71%/year from 2017 to 2021, ß = 0.979, P = 0.004. The number of new monoclonal prescriptions increased from 6 in 2007 to 111 in 2021. CONCLUSIONS: IBD incidence continues to increase in Southern England. Compounding prevalence and increased monoclonal usage has implications for service provision.


Subject(s)
COVID-19 , Colitis, Ulcerative , Crohn Disease , Inflammatory Bowel Diseases , Adolescent , Child , Chronic Disease , Colitis, Ulcerative/diagnosis , Colitis, Ulcerative/epidemiology , Crohn Disease/diagnosis , Crohn Disease/epidemiology , England/epidemiology , Female , Humans , Incidence , Inflammatory Bowel Diseases/epidemiology , Male , Prevalence , Prospective Studies
6.
Anal Chem ; 94(8): 3617-3628, 2022 03 01.
Article in English | MEDLINE | ID: mdl-35167252

ABSTRACT

Biofluids such as synovial fluid, blood plasma, and saliva contain several proteins which impart non-Newtonian properties to the biofluids. The concentration of such protein macromolecules in biofluids is regarded as an important biomarker for the diagnosis of several health conditions, including cardiovascular disorders, joint quality, and Alzheimer's. Existing technologies for the measurements of macromolecules in biofluids are limited; they require a long turnaround time, or require complex protocols, thus calling for alternative, more suitable, methodologies aimed at such measurements. According to the well-established relations for polymer solutions, the concentration of macromolecules in solutions can also be derived via measurement of rheological properties such as shear-viscosity and the longest relaxation time. We here introduce a microfluidic rheometer for rapid simultaneous measurement of shear viscosity and longest relaxation time of non-Newtonian solutions at different temperatures. At variance with previous technologies, our microfluidic rheometer provides a very short turnaround time of around 2 min or less thanks to the implementation of a machine-learning algorithm. We validated our platform on several aqueous solutions of poly(ethylene oxide). We also performed measurements on hyaluronic acid solutions in the clinical range for joint grade assessment. We observed monotonic behavior with the concentration for both rheological properties, thus speculating on their use as potential rheo-markers, i.e., rheological biomarkers, across several disease states.


Subject(s)
Machine Learning , Microfluidics , Microfluidics/methods , Rheology/methods , Solutions , Temperature , Viscosity
7.
Clin Exp Optom ; 105(1): 70-76, 2022 01.
Article in English | MEDLINE | ID: mdl-33730524

ABSTRACT

Clinical relevance: Mobility and fall risk may be important considerations in choosing between intraocular lenses.Background: Fall risk in older adults increases when wearing multifocal spectacles, but little is known about mobility among individuals with different types of intraocular lenses. This study compared visual function, fall risk and balance control following bilateral implantation of monofocal or multifocal intraocular lenses.Methods: This was a non-randomised, cross-sectional study involving adults with bilateral intraocular lenses. Participants completed questionnaires concerning physical functioning, fall history and balance-related confidence. Binocular visual acuity, contrast sensitivity (Pelli-Robson chart and computerized testing), depth perception and glare sensitivity were assessed. Physical performance measures included the Sensory Organization Test, preferred gait speed, Dynamic Gait Index and wayfinding in a virtual environment.Results: Fifteen participants (mean ± standard deviation, 67.1 ± 6.8 years) had monofocal intraocular lenses and 14 participants (68.1 ± 6.1 years) had multifocal intraocular lenses. Contrast sensitivity in the monofocal group was significantly better than that in the multifocal group (p = 0.02) at intermediate and high spatial frequencies. Contrast sensitivity of the monofocal group also was less affected by glare than the contrast sensitivity of the multifocal group, at an intermediate spatial frequency (p = 0.02). However, the multifocal group had significantly better Dynamic Gait Index scores (p = 0.04), even after controlling for perceived physical function.Conclusions: The participants with monofocal intraocular lenses generally had better contrast sensitivity than did those with multifocal intraocular lenses. However, the scores on a mobility test that is associated with fall risk were worse for those with monofocal lenses.


Subject(s)
Lens Implantation, Intraocular , Lenses, Intraocular , Aged , Contrast Sensitivity , Cross-Sectional Studies , Humans , Visual Acuity
8.
Mol Omics ; 18(1): 31-44, 2022 01 17.
Article in English | MEDLINE | ID: mdl-34709266

ABSTRACT

Lysine specific demethylase 1 (LSD1) regulates gene expression as part of the CoREST complex, along with co-repressor of REST (CoREST) and histone deacetylase 1 (HDAC1). CoREST is recruited to specific genomic loci by core components and numerous transient interactions with chromatin-associated factors and transcription factors. We hypothesise that many of these weaker and transient associations may be difficult to identify using traditional co-immunoprecipitation methods. We have therefore employed proximity-dependent biotin-identification (BioID) with four different members of the CoREST complex, in three different cell types, to identify a comprehensive network of LSD1/CoREST associated proteins. In HEK293T cells, we identified 302 CoREST-associated proteins. Among this group were 16 of 18 known CoREST components and numerous novel associations, including readers (CHD3, 4, 6, 7 and 8), writers (KMT2B and KMT2D) and erasers (KDM2B) of histone methylation. However, components of other HDAC1 containing complexes (e.g. Sin3) were largely absent. To examine the dynamic nature of the CoREST interactome in a primary cell type, we replaced endogenous LSD1 with BirA*-LSD1 in embryonic stem (ES) cells and performed BioID in pluripotent, early- and late-differentiating environments. We identified 156 LSD1-associated proteins of which 67 were constitutively associated across all three time-points (43%), including novel associations with the MMB and ChAHP complexes, implying that the majority of interactors are both dynamic and cell type dependent. In total, we have performed 16 independent BioID experiments for LSD1 in three different cell types, producing a definitive network of LSD1-assoicated proteins that should provide a major resource for the field.


Subject(s)
Biotin , Histone Demethylases , Cell Differentiation , Co-Repressor Proteins/genetics , Co-Repressor Proteins/metabolism , HEK293 Cells , Histone Demethylases/genetics , Histone Demethylases/metabolism , Humans , Nerve Tissue Proteins/genetics
9.
Arch Toxicol ; 95(9): 3101-3115, 2021 09.
Article in English | MEDLINE | ID: mdl-34245348

ABSTRACT

The in vitro micronucleus assay is a globally significant method for DNA damage quantification used for regulatory compound safety testing in addition to inter-individual monitoring of environmental, lifestyle and occupational factors. However, it relies on time-consuming and user-subjective manual scoring. Here we show that imaging flow cytometry and deep learning image classification represents a capable platform for automated, inter-laboratory operation. Images were captured for the cytokinesis-block micronucleus (CBMN) assay across three laboratories using methyl methanesulphonate (1.25-5.0 µg/mL) and/or carbendazim (0.8-1.6 µg/mL) exposures to TK6 cells. Human-scored image sets were assembled and used to train and test the classification abilities of the "DeepFlow" neural network in both intra- and inter-laboratory contexts. Harnessing image diversity across laboratories yielded a network able to score unseen data from an entirely new laboratory without any user configuration. Image classification accuracies of 98%, 95%, 82% and 85% were achieved for 'mononucleates', 'binucleates', 'mononucleates with MN' and 'binucleates with MN', respectively. Successful classifications of 'trinucleates' (90%) and 'tetranucleates' (88%) in addition to 'other or unscorable' phenotypes (96%) were also achieved. Attempts to classify extremely rare, tri- and tetranucleated cells with micronuclei into their own categories were less successful (≤ 57%). Benchmark dose analyses of human or automatically scored micronucleus frequency data yielded quantitation of the same equipotent concentration regardless of scoring method. We conclude that this automated approach offers significant potential to broaden the practical utility of the CBMN method across industry, research and clinical domains. We share our strategy using openly-accessible frameworks.


Subject(s)
Deep Learning , Flow Cytometry/methods , Micronucleus Tests/methods , Mutagens/toxicity , Automation, Laboratory , Benzimidazoles/administration & dosage , Benzimidazoles/toxicity , Carbamates/administration & dosage , Carbamates/toxicity , Cell Line , Cytokinesis/drug effects , DNA Damage/drug effects , Dose-Response Relationship, Drug , Humans , Methyl Methanesulfonate/administration & dosage , Methyl Methanesulfonate/toxicity , Mutagens/administration & dosage
10.
Nat Protoc ; 16(7): 3572-3595, 2021 07.
Article in English | MEDLINE | ID: mdl-34145434

ABSTRACT

Deep learning offers the potential to extract more than meets the eye from images captured by imaging flow cytometry. This protocol describes the application of deep learning to single-cell images to perform supervised cell classification and weakly supervised learning, using example data from an experiment exploring red blood cell morphology. We describe how to acquire and transform suitable input data as well as the steps required for deep learning training and inference using an open-source web-based application. All steps of the protocol are provided as open-source Python as well as MATLAB runtime scripts, through both command-line and graphic user interfaces. The protocol enables a flexible and friendly environment for morphological phenotyping using supervised and weakly supervised learning and the subsequent exploration of the deep learning features using multi-dimensional visualization tools. The protocol requires 40 h when training from scratch and 1 h when using a pre-trained model.


Subject(s)
Deep Learning , Image Cytometry/methods , Supervised Machine Learning , Software , User-Computer Interface
11.
Cytometry A ; 97(12): 1222-1237, 2020 12.
Article in English | MEDLINE | ID: mdl-32445278

ABSTRACT

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.


Subject(s)
Gastroenterology , Peyer's Patches , Animals , Flow Cytometry , Humans , Mice , Microscopy , Rats , Reproducibility of Results
12.
Arch Dis Child ; 105(7): 671-676, 2020 07.
Article in English | MEDLINE | ID: mdl-31937567

ABSTRACT

OBJECTIVE: Chronic diseases, such as inflammatory bowel disease (IBD), can impact negatively on education and social development. Examining the impact of IBD on school/college attendance for children and young people (CYP) is vital to provide targeted support to patients, families and schools. METHODS: We performed a cross-sectional survey to determine the school/college attendance rates, the reasons for absence related to IBD and facilitators or barriers to school/college attendance. In a subset of patients followed up locally, we performed a detailed review of hospital attendance data to assess healthcare burden. RESULTS: Two hundred and thirty-one questionnaires were given to CYP with IBD aged 5-17 years. Response rate was 74% (final sample 169). The median school/college attendance rate was 92.5%, significantly lower than all children in England (95.2%). 39.6% of children with IBD were persistently absent, defined nationally as missing 10% or more of school. Only five children (3%) had a 100% attendance record. Increasing age and use of monoclonal therapy were predictors of poor school attendance. Concerns about feeling unwell at school/college, access to toilets, keeping up with work and teachers' understanding of IBD are the main issues for CYP with IBD. There was a significant negative correlation between number of days in hospital and school attendance. CONCLUSION: IBD has a significant impact on school/college attendance, with hospital attendance, disease burden and school difficulties being major factors. Employing strategies to minimise healthcare burden and developing a partnership between health and education to support children with IBD will serve to facilitate school/college attendance.


Subject(s)
Antibodies, Monoclonal/therapeutic use , Education/statistics & numerical data , Inflammatory Bowel Diseases/epidemiology , Schools , Adolescent , Age Factors , Child , Child, Preschool , Comprehension , Cross-Sectional Studies , England/epidemiology , Faculty/psychology , Health Status , Humans , Inflammatory Bowel Diseases/drug therapy , Length of Stay/statistics & numerical data , Surveys and Questionnaires , Toilet Facilities
13.
Med Sci Sports Exerc ; 52(1): 259-266, 2020 01.
Article in English | MEDLINE | ID: mdl-31436733

ABSTRACT

PURPOSE: (i) To develop an automated measurement technique for the assessment of both the form and intensity of physical activity undertaken by children during play. (ii) To profile the varying activity across a cohort of children using a multivariate analysis of their movement patterns. METHODS: Ankle-worn accelerometers were used to record 40 min of activity during a school recess, for 24 children over five consecutive days. Activity events of 1.1 s duration were identified within the acceleration time trace and compared with a reference motif, consisting of a single walking stride acceleration trace, obtained on a treadmill operating at a speed of 4 km h. Dynamic time warping of motif and activity events provided metrics of comparative movement duration and intensity, which formed the data set for multivariate mapping of the cohort activity using a principal component analysis (PCA). RESULTS: The two-dimensional PCA plot provided clear differentiation of children displaying diverse activity profiles and clustering of those with similar movement patterns. The first component of the PCA correlated to the integrated intensity of movement over the 40-min period, whereas the second component informed on the temporal phasing of activity. CONCLUSIONS: By defining movement events and then quantifying them by reference to a motion-standard, meaningful assessment of highly varied activity within free play can be obtained. This allows detailed profiling of individual children's activity and provides an insight on social aspects of play through identification of matched activity time profiles for children participating in conjoined play.


Subject(s)
Child Behavior/physiology , Exercise/physiology , Movement/physiology , Play and Playthings , Accelerometry/instrumentation , Ankle , Child , Child, Preschool , Female , Humans , Male , Multivariate Analysis , Principal Component Analysis , Time and Motion Studies
14.
Hum Mov Sci ; 68: 102523, 2019 Dec.
Article in English | MEDLINE | ID: mdl-31683083

ABSTRACT

OBJECTIVE: While novel analytical methods have been used to examine movement behaviours, to date, no studies have examined whether a frequency-based measure, such a spectral purity, is useful in explaining key facets of human movement. The aim of this study was to investigate movement and gait quality, physical activity and motor competence using principal component analysis. METHODS: Sixty-five children (38 boys, 4.3 ±â€¯0.7y, 1.04 ±â€¯0.05 m, 17.8 ±â€¯3.2 kg, BMI; 16.2 ±â€¯1.9 kg∙m2) took part in this study. Measures included accelerometer-derived physical activity and movement quality (spectral purity), motor competence (Movement Assessment Battery for Children 2nd edition; MABC2), height, weight and waist circumference. All data were subjected to a principal component analysis, and the internal consistency of resultant components were assessed using Cronbach's alpha. RESULTS: Two principal components, with excellent internal consistency (Cronbach α >0.9) were found; the 1st principal component, termed "movement component", contained spectral purity, traffic light MABC2 score, fine motor% and gross motor% (α = 0.93); the 2nd principal component, termed "anthropometric component", contained weight, BMI, BMI% and body fat% (α = 0.91). CONCLUSION: The results of the present study demonstrate that accelerometric analyses can be used to assess motor competence in an automated manner, and that spectral purity is a meaningful, indicative, metric related to children's movement quality.


Subject(s)
Exercise/physiology , Motor Skills/physiology , Movement/physiology , Accelerometry/methods , Anthropometry/methods , Body Weight/physiology , Child , Child Development/physiology , Child, Preschool , Female , Gait/physiology , Humans , Male , Principal Component Analysis , Waist Circumference
15.
Transl Vis Sci Technol ; 8(3): 36, 2019 May.
Article in English | MEDLINE | ID: mdl-31211001

ABSTRACT

PURPOSE: To investigate the precision of visual fields (VFs) from semiautomated kinetic perimetry (SKP) on Octopus 900 perimeters, for children and adults with inherited retinal degenerations (IRDs). Goldmann manual kinetic perimetry has long been used in the diagnosis and follow-up of these patients, but SKP is becoming increasingly common. Octopus VFs (OVFs) and Goldmann VFs (GVFs) were both mapped on two occasions. METHODS: Nineteen females and 10 males with IRDs were tested on OVFs and GVFs, with two targets per test (V4e and one smaller target). Tests were performed in the same (randomized) order at two visits about 1 week apart. The VFs were digitized to derive isopter solid angles. Comparisons, within and between visits, were performed with paired t-tests and Bland-Altman plots. RESULTS: Median age was 20 years (range, 7-70; 10 participants aged ≤17 years old). There were no significant differences in solid angles between OVFs and GVFs (P ≥ 0.06) or between the two visits' solid angles on either perimeter (P ≥ 0.30). Between-visit test-retest variability for GVFs and OVFs was similar (P ≥ 0.73), with median values of approximately 9% to 13%. Overall variability was lower for children than adults (medians of 7.5% and 12.8%, respectively). CONCLUSIONS: Octopus SKP and Goldmann perimetry produced VFs of similar size and variability. TRANSLATIONAL RELEVANCE: Our study indicates that SKP provides a viable alternative to traditional Goldmann perimetry in clinical trials or care involving both children and adults with IRDs.

16.
Nat Commun ; 10(1): 2341, 2019 05 28.
Article in English | MEDLINE | ID: mdl-31138801

ABSTRACT

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.


Subject(s)
Endosomes/metabolism , Nanoparticles/metabolism , A549 Cells , Biological Transport , Cell Line , Endosomes/ultrastructure , High-Throughput Screening Assays , Humans , Image Processing, Computer-Assisted , Microscopy, Confocal , Nanoparticles/ultrastructure
17.
Essays Biochem ; 63(1): 97-107, 2019 04 23.
Article in English | MEDLINE | ID: mdl-30940741

ABSTRACT

Packaging the long and fragile genomes of eukaryotic species into nucleosomes is all well and good, but how do cells gain access to the DNA again after it has been bundled away? The solution, in every species from yeast to man, is to post-translationally modify histones, altering their chemical properties to either relax the chromatin, label it for remodelling or make it more compact still. Histones are subject to a myriad of modifications: acetylation, methylation, phosphorylation, ubiquitination etc. This review focuses on histone acylations, a diverse group of modifications which occur on the ε-amino group of Lysine residues and includes the well-characterised Lysine acetylation. Over the last 50 years, histone acetylation has been extensively characterised, with the discovery of histone acetyltransferases (HATs) and histone deacetylases (HDACs), and global mapping experiments, revealing an association of hyperacetylated histones with accessible, transcriptionally active chromatin. More recently, there has been an explosion in the number of unique short chain 'acylations' identified by MS, including: propionylation, butyrylation, crotonylation, succinylation, malonylation and 2-hydroxyisobutyrylation. These novel modifications add a range of chemical environments to histones, and similar to acetylation, appear to accumulate at transcriptional start sites and correlate with gene activity.


Subject(s)
Histones/metabolism , Nucleosomes/metabolism , Protein Processing, Post-Translational , Acetylation , Animals , DNA/genetics , DNA/metabolism , Gene Expression Regulation , Histones/chemistry , Histones/genetics , Humans , Lysine/chemistry , Nucleosomes/chemistry , Nucleosomes/genetics
18.
Doc Ophthalmol ; 136(3): 199-206, 2018 06.
Article in English | MEDLINE | ID: mdl-29934802

ABSTRACT

The International Society for Clinical Electrophysiology of Vision (ISCEV) standard for full-field electroretinography (ERG) describes a minimum procedure, but encourages more extensive testing. This ISCEV extended protocol describes an extension to the ERG standard, namely the photopic On-Off ERG, and outlines common clinical applications. A light stimulus duration of 150-200 ms is used in the presence of a rod-suppressing background to elicit cone-driven On- and Off-system ERG components. The On-response occurs after the stimulus onset and has a negative a-wave and positive b-wave. The Off d-wave is a positive component evoked by stimulus offset. Common diagnoses that may benefit from additional photopic On-Off ERG testing include retinal dystrophies and retinal disorders that cause dysfunction at a level that is post-phototransduction or post-receptoral. On-Off ERGs assess the relative involvement of On- and Off-systems and may be of use in the diagnosis of disorders such as complete and incomplete congenital stationary night blindness (complete and incomplete CSNB), melanoma-associated retinopathy, and some forms of autoimmune retinopathy. The photopic On-Off ERGs may also be useful in X-linked retinoschisis, Batten disease, Duchenne muscular dystrophy, spinocerebellar degeneration, quinine toxicity, and other retinal disorders.


Subject(s)
Color Vision/physiology , Electroretinography/standards , Retina/physiopathology , Retinal Dystrophies/physiopathology , Clinical Protocols/standards , Electrophysiology/standards , Humans , Photic Stimulation , Retinal Cone Photoreceptor Cells/physiology , Retinal Dystrophies/diagnosis , Societies, Medical/organization & administration
19.
Physiol Meas ; 39(4): 045007, 2018 04 26.
Article in English | MEDLINE | ID: mdl-29582781

ABSTRACT

OBJECTIVE: To quantify varied human motion and obtain an objective assessment of relative performance across a cohort. APPROACH: A wrist-worn magnetometer was used to record and quantify the complex motion patterns of 55 children aged 10 to 12 years old, generated during a fundamental movement skills programme. Sensor-based quantification of the physical activity used dynamic time warping of the magnetometer time series data for pairs of children. Pairwise comparison across the whole cohort produced a similarity matrix of all child to child correlations. Normative assessment scores were based on the Euclidean distance between n participants within an n - 1 multi-variate space, created from multi-dimensional scaling of the similarity matrix. The sensor-based scores were compared to the current standardised assessment which involves binary scoring of technique, outcome and time components by trained assessors. MAIN RESULTS: Visualisation of the relative performance using the first three axes of the multi-dimensional matrix, shows a 'performance sphere' in which children sit on concentric shells of increasing radius as performance deteriorates. Good agreement between standard and sensor scores is found, with Spearman rank correlation coefficients of the overall activity score in the range of 0.62-0.71 for different cohorts and a kappa statistic of 0.34 for categorisation of all 55 children into lower, middle, upper tertile and top 5% bands. SIGNIFICANCE: 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.


Subject(s)
Magnetometry/instrumentation , Movement , Child , Female , Humans , Time Factors
20.
J Mot Behav ; 50(5): 557-565, 2018.
Article in English | MEDLINE | ID: mdl-28985153

ABSTRACT

There is a dearth of suitable metrics capable of objectively quantifying motor competence. Further, objective movement quality characteristics during free play have not been investigated in pre-school children. The aims of this study were to characterize children's free play physical activity and investigate how gait quality characteristics cluster with free play in pre-school children (3-5 years old). Sixty-one children (39 boys; 4.3 ± 0.7 years, 1.04 ± 0.05 m, 17.8 ± 3.2 kg) completed the movement assessment battery for children and took part in free play while wearing an ankle- and hip-mounted accelerometer. Characteristics of movement quality were profiled using a clustering algorithm. Spearman's rho and the Mann-Whitney U tests were used to assess relationships between movement quality characteristics and motor competence classification differences in integrated acceleration and spectral purity, respectively. Significant differences were found between motor competency classifications for spectral purity and integrated acceleration (p < .001). Spectral purity was hierarchically clustered with motor competence and integrated acceleration. Significant positive correlations were found between spectral purity, integrated acceleration and motor competence (p < .001). This is the first study to report spectral purity in pre-school children and the results suggest that the underlying frequency component of movement is clustered with motor competence.


Subject(s)
Exercise/physiology , Gait/physiology , Movement/physiology , Acceleration , Accelerometry , Child, Preschool , Female , Humans , Male
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