Stabilizing the Proteomes of Acute Myeloid Leukemia Cells: Implications for Cancer Proteomics.

Previous work has shown that inhibition of abundant myeloid azurophil granule-associated serine proteases (ELANE [neutrophil elastase], PRTN3 [protease 3], and CTSG [Cathepsin G]) is required to stabilize some proteins in myeloid cells. We therefore hypothesized that effective inhibition of these proteases may be necessary for quantitative proteomic analysis of samples containing myeloid cells. To test this hypothesis, we thawed viably preserved acute myeloid leukemia cells from cryovials in the presence or the absence of diisopropyl fluorophosphate (DFP), a cell-permeable and irreversible serine protease inhibitor. Global proteomic analysis was performed, using label-free and isobaric peptide-labeling quantitation. The presence of DFP resulted in an increase of tryptic peptides (14-57%) and proteins (9-31%). In the absence of DFP, 11 to 31% of peptide intensity came from nontryptic peptides; 52 to 75% had cleavage specificity consistent with activities of ELANE-PRTN3. Treatment with DFP reduced the intensity of nontryptic peptides to 4-8% of the total. ELANE inhibition was 95%, based on diisopropyl phosphate modification of active site serine residue. Overall, the relative abundance of 20% of proteins was significantly altered by DFP treatment. These results suggest that active myeloid serine proteases, released during sample processing, can skew quantitative proteomic measurements. Finally, significant ELANE activity was also detected in Clinical Proteomics Tumor Analysis Consortium datasets of solid tumors (many of which have known myeloid infiltration). In the pancreatic cancer dataset, the median percentage of nontryptic intensity detected across patient samples was 34%, with many patient samples having more than half of their detected peptide intensity from nontryptic cleavage events consistent with ELANE-PRTN3 cleavage specificity. Our study suggests that in vitro cleavage of proteins by myeloid serine proteases may be relevant for proteomic studies of any tumor that contains infiltrating myeloid cells.

First Real-World Experience With Bigfoot Unity: A 6-Month Retrospective Analysis.

The Bigfoot Unity Diabetes Management System, a smart pen cap system cleared by the U.S. Food and Drug Administration in May 2021, incorporates continuous glucose monitoring data, real-time glycemic alerts, and clinician-directed dose recommendations. This study analyzed real-world clinical outcomes data for an initial cohort (n = 58, from 13 clinics) managing multiple daily injection insulin therapy using the pen cap system for 6 months. We examined glycemic control, including hypoglycemia events and interaction with and use of the pen cap system. In a cohort mainly consisting of adults with type 2 diabetes and an average age of 62 years, the results demonstrate close adherence to established glycemic targets, including a relatively short amount of time spent in the hypoglycemic range.

Intra-unit reliability and movement variability of submission grappling external load as measured by torso mounted accelerometery.

Submission grappling consists of skills and movements used in combat sports to physically control opponents whilst trying to apply choke holds and joint locks. There is currently no accepted method of monitoring external load in grappling-based sports due to the absence of key variables such as distance, velocity or time. The primary aim of this study was to determine whether PlayerLoad is a reliable variable for measuring external load of submission grappling movements, with a secondary aim of determining the between repetition variance of submission grappling movements. 7 experienced submission grapplers were recruited. Each wore a torso mounted Catapult® Optimeye S5 microelectromechanical systems (MEMS) device and completed 5 repetitions of each of the following: 4 submission techniques; 5 transition techniques; 2 guard pass techniques; 2 takedown techniques. Accumulated PlayerLoad (PLdACC) was recorded as a marker of absolute load, with accumulated PlayerLoad per minute (PLdACC∙min-1) representing relative load. Reliability of each was assessed using intraclass correlation coefficient (ICC(3,1)) (≥ 0.70). Between repetition movement variation was assessed via coefficient of variation with 95% confidence intervals (CV, 95%CI) (acceptable ≤ 15%, good ≤ 10%). PLdACC ICC(3,1) range = 0.78-0.98, with CV range = 9-22%. PLdACC∙min-1 ICC(3,1) range = 0.83-0.98, with CV range = 11-19%. Though several variables displayed CV > 15%, all had 95%CI lower boundaries ≤ 15%. Whilst PlayerLoad was found to be a reliable measure for submission grappling, relatively high CVs across most techniques examined suggest PlayerLoad may not be appropriate for measuring changes in external load for individual movements in submission grappling. However, it may prove a useful tool for monitoring the external load of full, grappling-based, training sessions within an individual.

Cold for centuries: a brief history of cryotherapies to improve health, injury and post-exercise recovery.

For centuries, cold temperatures have been used by humans for therapeutic, health and sporting recovery purposes. This application of cold for therapeutic purposes is regularly referred to as cryotherapy. Cryotherapies including ice, cold-water and cold air have been popularised by an ability to remove heat, reduce core and tissue temperatures, and alter blood flow in humans. The resulting downstream effects upon human physiologies providing benefits that include a reduced perception of pain, or analgesia, and an improved sensation of well-being. Ultimately, such benefits have been translated into therapies that may assist in improving post-exercise recovery, with further investigations assessing the role that cryotherapies can play in attenuating the ensuing post-exercise inflammatory response. Whilst considerable progress has been made in our understanding of the mechanistic changes associated with adopting cryotherapies, research focus tends to look towards the future rather than to the past. It has been suggested that this might be due to the notion of progress being defined as change over time from lower to higher states of knowledge. However, a historical perspective, studying a subject in light of its earliest phase and subsequent evolution, could help sharpen one's vision of the present; helping to generate new research questions as well as look at old questions in new ways. Therefore, the aim of this brief historical perspective is to highlight the origins of the many arms of this popular recovery and treatment technique, whilst further assessing the changing face of cryotherapy. We conclude by discussing what lies ahead in the future for cold-application techniques.

Effect of alterations in whole-body cryotherapy (WBC) exposure on post-match recovery markers in elite Premier League soccer players.

The use of whole-body cryotherapy (WBC) as a recovery intervention is prevalent amongst elite soccer players. However, there is a distinct lack of data available around chronic WBC use and post-match recovery markers in elite soccer. The aim of this study was to investigate the impact of different levels of WBC exposure on subjective and objective measures of post-match recovery in elite soccer players during a chronic exposure period. Sixteen male senior professional outfield soccer players participated in this study over two seasons. K means cluster analysis was used to classify low (-114 ± 2°C for 133 ± 2 s), medium (-121 ± 1°C for 173 ± 2 s) and high (-133 ± 1°C for 181 ± 2 s) cryotherapy exposure indexes (CEI). Salivary markers (immunoglobulin A (IgA) and alpha amylase (AA)) and subjective wellness scores (perceived fatigue, sleep quality, general muscle soreness and stress) were collected post-match across both seasons. Training load (session-RPE) was collected and used as a covariate to control for the load amongst groups. No differences were seen in perceived measures of wellness and salivary AA. Significantly lower IgA concentrations were observed in the medium CEI group (255 ± 32 µg∙ml-1) compared to the low (328 ± 38 µg∙ml-1) and high (306 ± 32 µg∙ml-1) groups. Therefore, increasing the level of chronic WBC exposure appears to have no additional benefit on subjective recovery and alpha amylase response post-match. However, there appears to be an optimal chronic WBC dose with regards to IgA response.

Exogenous carbohydrate and regulation of muscle carbohydrate utilisation during exercise.

PURPOSE: Carbohydrates (CHO) are one of the fundamental energy sources during prolonged steady state and intermittent exercise. The consumption of exogenous CHO during exercise is common place, with the aim to enhance sporting performance. Despite the popularity around exogenous CHO use, the process by which CHO is regulated from intake to its use in the working muscle is still not fully appreciated. Recent studies utilizing the hyperglycaemic glucose clamp technique have shed light on some of the potential barriers to CHO utilisation during exercise. The present review addresses the role of exogenous CHO utilisation during exercise, with a focus on potential mechanisms involved, from glucose uptake to glucose delivery and oxidation at the different stages of regulation. METHODS: Narrative review. RESULTS: A number of potential barriers were identified, including gastric emptying, intestinal absorption, blood flow (splanchnic and muscle), muscle uptake and oxidation. The relocation of glucose transporters plays a key role in the regulation of CHO, particularly in epithelial cells and subsequent transport into the blood. Limitations are also apparent when CHO is infused, particularly with regards to blood flow and uptake within the muscle. CONCLUSION: We highlight a number of potential barriers involved with the regulation of both ingested and infused CHO during exercise. Future work on the influence of longitudinal training within the regulation processes (such as the gut) is warranted to further understand the optimal type, dose and method of CHO delivery to enhance sporting performance.

Identifiers for the 21st century: How to design, provision, and reuse persistent identifiers to maximize utility and impact of life science data.

In many disciplines, data are highly decentralized across thousands of online databases (repositories, registries, and knowledgebases). Wringing value from such databases depends on the discipline of data science and on the humble bricks and mortar that make integration possible; identifiers are a core component of this integration infrastructure. Drawing on our experience and on work by other groups, we outline 10 lessons we have learned about the identifier qualities and best practices that facilitate large-scale data integration. Specifically, we propose actions that identifier practitioners (database providers) should take in the design, provision and reuse of identifiers. We also outline the important considerations for those referencing identifiers in various circumstances, including by authors and data generators. While the importance and relevance of each lesson will vary by context, there is a need for increased awareness about how to avoid and manage common identifier problems, especially those related to persistence and web-accessibility/resolvability. We focus strongly on web-based identifiers in the life sciences; however, the principles are broadly relevant to other disciplines.

Human regular U-500 insulin via continuous subcutaneous insulin infusion versus multiple daily injections in adults with type 2 diabetes: The VIVID study.

AIM: To compare the safety and efficacy of U500-R delivered by a novel, specifically designed U500-R insulin pump with U-500R delivered by multiple daily injections (MDI). METHODS: The phase 3 VIVID study randomized people with type 2 diabetes to U-500R by continuous subcutaneous insulin infusion (CSII) or MDI. Participants (aged 18-85 years) had HbA1c ≥7.5% and ≤12.0% and a total daily dose of insulin >200 and ≤600 U/day. After a 2-week transition to three times daily injections of U-500R, participants were treated for 24 weeks with U-500R by CSII or MDI. Treatment arms were compared using mixed model repeated measures analysis. RESULTS: The study randomized 420 participants (CSII: 209, MDI: 211) with 365 completers. Mean changes from baseline were: HbA1c, -1.27% (-13.9 mmol/mol) with CSII and -0.85% (-9.3 mmol/mol) with MDI (difference - 0.42% [-4.6 mmol/mol], P <0.001); fasting plasma glucose, -33.9 mg/dL (-1.9 mmol/L) with CSII and 1.7 mg/dL (0.09 mmol/L) with MDI (difference - 35.6 mg/dL [-2.0 mmol/L], P <0.001); total daily dose, 2.8 U with CSII and 51.3 U with MDI (P < 0.001). Weight changes and rates of documented symptomatic and severe hypoglycaemia were similar between groups; the CSII group had a higher rate of nocturnal hypoglycaemia. CONCLUSIONS: In type 2 diabetes requiring high doses of insulin, both methods of U-500R delivery lowered HbA1c. However, the CSII group attained greater HbA1c reduction with significantly less insulin. Individualized dose titration will be important to balance glycaemic control with hypoglycaemia risk.

A 3-day dietary manipulation affects muscle glycogen and results in modifications of carbohydrate and fat metabolism during exercise when hyperglycaemic.

PURPOSE: The effect of hyperglycaemia on exercise with low and elevated muscle glycogen on glucose utilization (GUR), carbohydrate and fat oxidation, hormonal and metabolite responses, as well as rating of perceived exertion (RPE) were explored. METHODS: Five healthy trained males were exercised for 90 min at 70% V̇O2max in two trials, while glucose was infused intravenously at rates to "clamp" blood glucose at 12 mM. On one occasion, participants were 'loaded' with carbohydrate (CHO-L), whilst on a separate occasion, participants were glycogen depleted (CHO-D). Prior exercise and dietary manipulations produced the 'loaded' and 'depleted' states. RESULTS: The CHO-L and CHO-D conditions resulted in muscle glycogen concentrations of 377 and 159 mmol/g dw, respectively. Hyperglycaemia elevated plasma insulin concentrations with higher levels for CHO-L than for CHO-D (P < 0.01). Conversely, CHO-D elevated plasma adrenaline and noradrenaline higher than CHO-L (P < 0.05). Plasma fat metabolites (NEFA, ß-hydroxybutyrate, and glycerol) were higher under CHO-D than CHO-L (P < 0.01). The resultant was that the rates of total carbohydrate and fat oxidation were elevated and depressed for loaded CHO-L vs CHO-D respectively (P < 0.01), although no difference was found for GUR (P > 0.05). The RPE over the exercise period was higher for CHO-D than CHO-L (P < 0.05). CONCLUSION: Hyperglycaemia during exercise, when muscle glycogen is reduced, attenuates insulin but promotes catecholamines and fat metabolites. The effect is a subsequent elevation of fat oxidation, a reduction in CHO oxidation without a concomitant increase in GUR, and an increase in RPE.

Hyperinsulinaemia and hyperglycaemia promote glucose utilization and storage during low- and high-intensity exercise.

PURPOSE: The effect of hyperglycaemia with and without additional insulin was explored at a low and high intensity of exercise (40% vs 70% VO2peak) on glucose utilization (GUR), carbohydrate oxidation, non-oxidative glucose disposal (NOGD), and muscle glycogen. METHODS: Eight healthy trained males were exercised for 120 min in four trials, twice at 40% VO2peak and twice at 70% VO2peak, while glucose was infused intravenously (40%G; 70%G) at rates to "clamp" blood glucose at 10 mM. On one occasion at each exercise intensity, insulin was also infused at 40 mU/m2/per min (i.e. 40%GI and 70%GI). The glucose and insulin infusion began 30 min prior to exercise and throughout exercise. A muscle biopsy was taken at the end of exercise for glycogen analysis. RESULTS: Hyperglycaemia significantly elevated plasma insulin concentration (p < 0.001), although no difference was observed between the exercise intensities. Insulin infusion during both mild and severe exercise resulted in increased insulin concentrations (p < 0.01) and GUR (p < 0.01) compared with glucose (40%GI by 25.2%; 70%GI by 26.2%), but failed to significantly affect carbohydrate, fat and protein oxidation. NOGD was significantly higher for GI trials at both intensities (p < 0.05) with storage occurring during both lower intensities (62.7 ± 19.6 g 40%GI; 127 ± 20.7 g 40%GI) and 70%GI (29.0 ± 20.0 g). Muscle glycogen concentrations were significantly depleted from rest (p < 0.01) after all four trials. CONCLUSION: Hyperinsulinaemia in the presence of hyperglycaemia during both low- and high-intensity exercise promotes GUR and NOGD, but does not significantly affect substrate oxidation.

Workload and Injury in Professional Soccer Players: Role of Injury Tissue Type and Injury Severity.

The purpose of the present study was to examine the influence of workload prior to injury on injury (tissue type and severity) in professional soccer players. Twenty-eight days of retrospective training data prior to non-contact injuries (n=264) were collated from 192 professional soccer players. Each injury tissue type (muscle, tendon and ligament) and severity (days missed) were categorised by medical staff. Training data were recorded using global positioning system (GPS) devices for total distance (TD), high speed distance (HSD,>5.5 m/s-1), and sprint distance (SPR,>7.0 m/s-1). Accumulated 1, 2, 3, 4-weekly loads and acute:chronic workload ratios (ACWR) (coupled, uncoupled and exponentially weighted moving average (EWMA) approaches) were calculated. Workload variables and injury tissue type were compared using a one-way ANOVA. The association between workload variables and injury severity were examined using a bivariate correlation. There were no differences in accumulated weekly loads and ACWR calculations between muscle, ligament, and tendon injuries (P>0.05). Correlations between each workload variable and injury severity highlighted no significant associations (P>0.05). The present findings suggest that the ability of accumulated weekly workload or ACWR methods to differentiate between injury type and injury severity are limited using the present variables.

Open Targets: a platform for therapeutic target identification and validation.

We have designed and developed a data integration and visualization platform that provides evidence about the association of known and potential drug targets with diseases. The platform is designed to support identification and prioritization of biological targets for follow-up. Each drug target is linked to a disease using integrated genome-wide data from a broad range of data sources. The platform provides either a target-centric workflow to identify diseases that may be associated with a specific target, or a disease-centric workflow to identify targets that may be associated with a specific disease. Users can easily transition between these target- and disease-centric workflows. The Open Targets Validation Platform is accessible at https://www.targetvalidation.org.

Carbohydrate oxidation and glucose utilisation under hyperglycaemia in aged and young males during exercise at the same relative exercise intensity.

PURPOSE: The purpose of the present study was to investigate the age-related carbohydrate oxidation and glucose utilisation rate response during exercise at the same relative intensity under hyperglycaemia in aged and young males. METHODS: 16 endurance-trained aged (n = 8; 69.1 ± 5.2 year) and young (n = 8; 22.4 ± 2.9 year) males were studied during 40 min of cycling exercise (60% [Formula: see text]) under both hyperglycaemic and euglycaemic (control) conditions. Venous blood samples were collected at baseline, post-infusion, mid- and post-exercise. Carbohydrate and fat oxidation rates were determined at both 15 and 35 min during exercise, and glucose utilisation rates were calculated. RESULTS: The aged group displayed significantly lower rates of carbohydrate oxidation during exercise during maintained hyperglycemia (15 min = 2.3 ± 0.4 vs. 1.6 ± 0.5 g min-1; 35 min = 2.3 ± 0.5 vs. 1.5 ± 0.5 g min-1) and control (15 min = 2.2 ± 0.4 vs. 1.6 ± 0.7 g min-1; 35 min = 1.9 ± 0.7 vs. 1.3 ± 0.7 g min-1) conditions (P = 0.01). The rate of glucose utilisation during exercise was also significantly reduced (85.76 ± 23.95 vs. 56.67 ± 15.09 µM kg-1 min-1). There were no differences between age groups for anthropometric measures, fat oxidation, insulin, glucose, NEFA, glycerol and lactate (P > 0.05) although hyperglycemia resulted in elevated glucose and insulin, and attenuated fat metabolite levels. CONCLUSION: Our findings highlight that ageing results in a reduction in carbohydrate oxidation and utilisation rates during exercise at the same relative exercise intensity.

Use of clinical quality indicators to improve lung cancer care in a regional/rural network of health services.

PROBLEM: Optimal lung cancer care requires multidisciplinary team input, with access to specialised diagnostic and therapeutic services that may be limited in rural or regional areas and impact clinical outcomes. Clinical quality indicators can be used to measure the quality of care delivered to patients with lung cancer in a region and identify areas for improvement. We describe the implementation of internationally recognised clinical quality indicators for lung cancer care in the Barwon South Western region. DESIGN: The consensus of an expert panel was used for the selection of clinical quality indicators. The data were retrospectively collected from the Evaluation of Cancer Outcomes Barwon South West Registry, which systematically records detailed information on all new patients with cancer in the region. SETTING: Region-based health service. KEY MEASURES FOR IMPROVEMENT: Adherence to clinical quality indicator targets. STRATEGIES FOR CHANGE: Clinical quality indicators, which fall short of the expected targets, highlight areas for improvement in the service provided to patients with lung cancer. These results have prompted changes in the service offered to these patients, such as the introduction of a multidisciplinary lung cancer clinic. EFFECTS OF CHANGE: The multidisciplinary lung cancer clinic has streamlined the access to lung cancer services, including specialist consultations, diagnostics and therapeutic services, in a regional setting. Ongoing data collection is required to determine the effect of such changes on adherence to clinical quality indicator targets. LESSONS LEARNT: The regular monitoring of clinical quality indicators serves as a useful method of quality assurance in the care of patients with lung cancer. We expect these clinical quality indicators to also be used by other health services to analyse and improve services provided to patients with lung cancer.

Disease Ontology 2015 update: an expanded and updated database of human diseases for linking biomedical knowledge through disease data.

The current version of the Human Disease Ontology (DO) (http://www.disease-ontology.org) database expands the utility of the ontology for the examination and comparison of genetic variation, phenotype, protein, drug and epitope data through the lens of human disease. DO is a biomedical resource of standardized common and rare disease concepts with stable identifiers organized by disease etiology. The content of DO has had 192 revisions since 2012, including the addition of 760 terms. Thirty-two percent of all terms now include definitions. DO has expanded the number and diversity of research communities and community members by 50+ during the past two years. These community members actively submit term requests, coordinate biomedical resource disease representation and provide expert curation guidance. Since the DO 2012 NAR paper, there have been hundreds of term requests and a steady increase in the number of DO listserv members, twitter followers and DO website usage. DO is moving to a multi-editor model utilizing Protégé to curate DO in web ontology language. This will enable closer collaboration with the Human Phenotype Ontology, EBI's Ontology Working Group, Mouse Genome Informatics and the Monarch Initiative among others, and enhance DO's current asserted view and multiple inferred views through reasoning.

Sport science internships for learning: a critical view.

Sport science has gained vast popularity with students who have an interest in both physiology and the underpinning mechanisms of exercise concerning performance and health. The high numbers of graduates each year, coupled with the low number of graduate positions working in sports, has led to a high level of competition between students. To stand out from the crowd, sport science students may undertake an internship placement as part of their course, designed to enhance theoretical, practical, and soft skills in an applied setting. In the present article, we highlight some of the positives and negatives of sport science internships and ways in which they can be implemented and facilitated. Suggestions have also been provided to make students more aware of the reality of working in professional sports, which includes awareness of the potential for long and unsociable hours of work.

Expression Atlas update--a database of gene and transcript expression from microarray- and sequencing-based functional genomics experiments.

Expression Atlas (http://www.ebi.ac.uk/gxa) is a value-added database providing information about gene, protein and splice variant expression in different cell types, organism parts, developmental stages, diseases and other biological and experimental conditions. The database consists of selected high-quality microarray and RNA-sequencing experiments from ArrayExpress that have been manually curated, annotated with Experimental Factor Ontology terms and processed using standardized microarray and RNA-sequencing analysis methods. The new version of Expression Atlas introduces the concept of 'baseline' expression, i.e. gene and splice variant abundance levels in healthy or untreated conditions, such as tissues or cell types. Differential gene expression data benefit from an in-depth curation of experimental intent, resulting in biologically meaningful 'contrasts', i.e. instances of differential pairwise comparisons between two sets of biological replicates. Other novel aspects of Expression Atlas are its strict quality control of raw experimental data, up-to-date RNA-sequencing analysis methods, expression data at the level of gene sets, as well as genes and a more powerful search interface designed to maximize the biological value provided to the user.

The EBI RDF platform: linked open data for the life sciences.

MOTIVATION: Resource description framework (RDF) is an emerging technology for describing, publishing and linking life science data. As a major provider of bioinformatics data and services, the European Bioinformatics Institute (EBI) is committed to making data readily accessible to the community in ways that meet existing demand. The EBI RDF platform has been developed to meet an increasing demand to coordinate RDF activities across the institute and provides a new entry point to querying and exploring integrated resources available at the EBI.

ArrayExpress update--trends in database growth and links to data analysis tools.

The ArrayExpress Archive of Functional Genomics Data (http://www.ebi.ac.uk/arrayexpress) is one of three international functional genomics public data repositories, alongside the Gene Expression Omnibus at NCBI and the DDBJ Omics Archive, supporting peer-reviewed publications. It accepts data generated by sequencing or array-based technologies and currently contains data from almost a million assays, from over 30 000 experiments. The proportion of sequencing-based submissions has grown significantly over the last 2 years and has reached, in 2012, 15% of all new data. All data are available from ArrayExpress in MAGE-TAB format, which allows robust linking to data analysis and visualization tools, including Bioconductor and GenomeSpace. Additionally, R objects, for microarray data, and binary alignment format files, for sequencing data, have been generated for a significant proportion of ArrayExpress data.