Functional assays reflective of cancer hallmarks in BT-549 cells are not impacted by media supplemented with exercise-trained plasma.

NEW FINDINGS: What is the central question of this study? Little is known regarding the effects of media supplemented with resting plasma from exercise-trained individuals, despite the established bioactive effects of acutely exercised samples. Does media supplemented with resting plasma from endurance-trained, strength-trained or recreationally active controls impact hallmarks of cancer in BT-549 cells? What is the main finding and its importance? Supplementing media with plasma from these trained athletes did not impact proliferation, migration, invasion or anoikis resistance compared to plasma from recreationally-active controls. These findings suggest that 'anti-cancer' effects of exercise are not present in resting blood samples of exercise-trained individuals. ABSTRACT: Media supplemented with sera from acutely exercised men has been shown to have 'anti-cancer' effects on prostate and breast cancer cell lines. This study investigated whether media supplemented with plasma samples taken at rest (≥30 h since the most recent exercise session) from men who were endurance-trained (END), strength-trained (STR) or recreationally active controls (CON) impacted the results of four assays that mimic hallmarks of cancer (proliferation, migration, extracellular matrix invasion and anoikis resistance) in the BT-549 breast cancer cell line. Compared to control conditions of either serum-free media or fetal bovine serum as appropriate, BT-549 cells cultured with plasma-supplemented media regardless of group resulted in greater cell proliferation (∼20-50%) and cell migration (∼15-20%), and lower extracellular matrix invasion (∼10-20%) and anoikis resistance (∼15-20%). Supplementing media with plasma from END or STR did not impact any outcomes of these assays compared to plasma from CON. Media supplemented with human plasma can impact functional assays reflective of cancer hallmarks in BT-549 cells, but effects of exercise on proliferation, migration, extracellular matrix invasion and anoikis resistance were not evident in resting blood samples of individuals with a prior history of exercise training.

The separation and identification of circulating small extracellular vesicles from endurance-trained, strength-trained and recreationally active men.

Small extracellular vesicles (EV) are membrane-encapsulated particles that carry bioactive cargoes, are released by all cell types and are present in all human biofluids. Changes in EV profiles and abundance occur in response to acute exercise, but this study investigated whether individuals with divergent histories of exercise training (recreationally active controls - CON; endurance-trained - END; strength-trained - STR) presented with varied abundances of small EVs in resting samples and whether the abundance of small EVs differed within each group across two measurement days. Participants (n = 38, all male; CON n = 12, END n = 13, STR n = 13) arrived at the lab on two separate occasions in a rested, overnight fasted state, with standardisation of time of day of sampling, recent dietary intake, time since last meal and time since last exercise training session (∼40 h). Whole blood samples were collected and separated into plasma from which small EVs were separated using size exclusion chromatography and identified in accordance with the Minimal Information For Studies of Extracellular Vesicles (MISEV) guidelines. No differences in the abundance of small EVs were observed within or between groups across multiple methods of small EV identification (nanoparticle tracking analysis, flow cytometry, immunoblot of specific EV markers). Targeted metabolomics of the small EV preparations identified 96 metabolites that were associated with the structure and function of small EVs, with no statistically significant differences in concentrations observed across groups. The results of the current study suggest that the abundance and metabolomic profile of small EVs derived from men with divergent histories of exercise training are similar to those in resting blood samples. KEY POINTS: Extracellular vesicles (EV) are membrane-encapsulated particles that are present in circulation and carry bioactive materials as 'cargo'. The abundance and profile of small EVs are responsive to acute exercise, but little is known about the relationship between small EVs and exercise training. This study examined the abundance, and a targeted metabolomic profile, of small EVs separated from the blood of endurance athletes, strength athletes and recreationally active controls at rest (∼40 h after the most recent exercise session) on two separate but identical lab visits. No differences were observed in the abundance or metabolomic profile of small EV preparations between the groups or between the lab visits within each group. Further research should determine whether the bioactive cargoes (e.g. RNA, protein and additional metabolites) carried within EVs are altered in individuals with divergent histories of exercise training or in response to exercise training interventions.

Within-Subject Variability and the Influence of Exercise Training History on the Resting Plasma Metabolome in Men.

This study investigated within-subject variability in the circulating metabolome under controlled conditions, and whether divergent exercise training backgrounds were associated with alterations in the circulating metabolome assessed in resting samples. Thirty-seven men comprising of endurance athletes (END; body mass, 71.0 ± 6.8 kg; fat-free mass index, 16.9 ± 1.1 kg/m2), strength athletes (STR; 94.5 ± 8.8 kg; 23.0 ± 1.8 kg/m2), and recreationally active controls (CON; 77.6 ± 7.7 kg; 18.1 ± 1.0 kg/m2) provided blood samples after an overnight fast on two separate occasions controlled for time of day of sampling, recent dietary intake, time since last meal, and time since last exercise training session. A targeted profile of metabolites, performed using liquid chromatography and mass spectrometry on plasma samples, identified 166 individual metabolites and metabolite features, which were analyzed with intraclass correlation coefficients, a multilevel principal component analysis, and univariate t tests adjusted for multiple comparisons. The median intraclass correlation coefficient was .49, with 46 metabolites displaying good reliability and 31 metabolites displaying excellent reliability. No difference in the abundance of any individual metabolite was identified within groups when compared between visits, but a combined total of 44 metabolites were significantly different (false discovery rate <0.05) between groups (END vs. CON, 42 metabolites; STR vs. CON, 10 metabolites; and END vs. STR, five metabolites). Under similar measurement conditions, the reliability of resting plasma metabolite concentrations varies largely at the level of individual metabolites with ∼48% of metabolites displaying good-to-excellent reliability. However, a history of exercise training was associated with alterations in the abundance of ∼28% of metabolites in the targeted profile employed in this study.

The resting serum metabolome in response to short-term sprint interval training.

PURPOSE: To investigate the response of a targeted fraction of (168 metabolites) of the resting serum metabolome to 9 sessions of sprint interval training (SIT). METHODS: Thirty-four recreationally active males provided resting blood samples before (baseline) and 48-72 h after (post) a short-term (9 sessions) cycle ergometer-based SIT intervention. A targeted analysis of 168 metabolites was performed on serum using liquid chromatography mass spectrometry (LC-MS). 160 distinct metabolites were identified and combined with 4 calculated metabolite sums and 3 calculated metabolite ratios creating a panel of 167 individual factors. Data were analysed using principal component analysis and univariate testing of all factors classified into 5 metabolite subgroups. RESULTS: SIT improved anaerobic capacity measured by average power output during a Wingate test (p < 0.01; mean difference = 38 W, 95% confidence interval [26, 51]) and aerobic capacity measured by average power output in a 20 min cycling test (p < 0.01; 17 W [12, 23]). Limited separation was discernible in the targeted serum metabolome between baseline and post-intervention when projected on the first and second principal component(s). However, univariate testing identified 11 fatty acids that had lower concentrations (false discovery rate < 0.05) in post-intervention samples. CONCLUSIONS: These findings demonstrate that this short-term SIT intervention had limited effect on the serum metabolome at rest, but a subfraction of fatty acids are potentially sensitive to short-term exercise training.

Exercise Training and Circulating Small Extracellular Vesicles: Appraisal of Methodological Approaches and Current Knowledge.

In response to acute exercise, an array of metabolites, nucleic acids, and proteins are enriched in circulation. Collectively termed "exercise factors," these molecules represent a topical area of research given their speculated contribution to both acute exercise metabolism and adaptation to exercise training. In addition to acute changes induced by exercise, the resting profile of circulating exercise factors may be altered by exercise training. Many exercise factors are speculated to be transported in circulation as the cargo of extracellular vesicles (EVs), and in particular, a sub-category termed "small EVs." This review describes an overview of exercise factors, small EVs and the effects of exercise, but is specifically focused on a critical appraisal of methodological approaches and current knowledge in the context of changes in the resting profile small EVs induced by exercise training, and the potential bioactivities of preparations of these "exercise-trained" small EVs. Research to date can only be considered preliminary, with interpretation of many studies hindered by limited evidence for the rigorous identification of small EVs, and the conflation of acute and chronic responses to exercise due to sample timing in proximity to exercise. Further research that places a greater emphasis on the rigorous identification of small EVs, and interrogation of potential bioactivity is required to establish more detailed descriptions of the response of small EVs to exercise training, and consequent effects on exercise adaptation.

Habitual Minimalist Shod Running Biomechanics and the Acute Response to Running Barefoot.

The aim of the study was to determine whether habitual minimalist shoe runners present with purported favorable running biomechanithat reduce running injury risk such as initial loading rate. Eighteen minimalist and 16 traditionally cushioned shod runners were assessed when running both in their preferred training shoe and barefoot. Ankle and knee joint kinetics and kinematics, initial rate of loading, and footstrike angle were measured. Sagittal ankle and knee joint stiffness were also calculated. Results of a two-factor ANOVA presented no group difference in initial rate of loading when participants were running either shod or barefoot; however, initial loading rate increased for both groups when running barefoot (p=0.008). Differences in footstrike angle were observed between groups when running shod, but not when barefoot (minimalist:8.71±8.99 vs. traditional: 17.32±11.48 degrees, p=0.002). Lower ankle joint stiffness was found in both groups when running barefoot (p=0.025). These findings illustrate that risk factors for injury potentially differ between the two groups. Shoe construction differences do change mechanical demands, however, once habituated to the demands of a given shoe condition, certain acute favorable or unfavorable responses may be moderated. The purported benefits of minimalist running shoes in mimicking habitual barefoot running is questioned, and risk of injury may not be attenuated.