Sixty subjects were tested five times per waking day on two occasions for accuracy and reliability in throwing 20 darts at a target. Two experimental conditions were investigated: following a normal nocturnal sleep (7–8 h sleep, normal) and after having retired to bed 4 h later than normal the previous night but rising at the normal time (3–4 h sleep, sleep deprivation). Sublingual (core) temperature and subjective estimates of alertness and fatigue were measured in all sessions. Performance at throwing darts was assessed by three methods: mean distance of the dart from the bulls-eye; number of times the target was missed; and variability of the scores from the darts thrown. There was no evidence that performance was affected by physical fatigue arising during the course of throwing the 20 darts. All variables showed significant diurnal rhythms, those of alertness and performance being phased over 1 h earlier than core temperature, and that of fatigue over 1 h earlier than the inverse of temperature. Core temperature was not affected by sleep deprivation, but all other variables showed significant changes, indicative of mood and performance decrement. Increasing time awake was associated with decreased alertness and increased fatigue, as well as slight negative effects upon performance. We conclude that the simple task of throwing darts at a target provides information about chronobiological changes in circumstances where time awake and sleep loss might affect psychomotor performance. (Author correspondence: b.j.edwards@ljmu.ac.uk)

We have investigated the effects that sleep restriction (3-h sleep during two consecutive nights) have on an evening (17:00 h) submaximal weightlifting session; and whether this performance improves following a 1-h post-lunch powernap. Fifteen resistance-trained males participated in this study. Before the experimental protocol commenced, 1RM bench press and inclined leg press and normative habitual sleep were recorded. Participants were familiarised with the testing protocol, then completed three experimental conditions with two nights of prescribed sleep: (i) Normal (N): retire at 23:00 h and wake at 06:30 h, (ii) partial sleep-deprivation (SD): retire at 03:30 h and wake at 06:30 h and (iii) partial sleep-deprivation with nap (SD_N): retire at 03:30 h and wake at 06:30 h with a 1-h nap at 13:00 h. Each condition was separated by at least 7 days and the order of administration was randomised and counterbalanced. Rectal (T_rec) and mean skin (T_s) temperatures, Profile of Mood Scores, subjective tiredness, alertness and sleepiness values were measured at 08:00, 11:00, 14:00 and 17:00 h on the day of the weightlifting session. Following the final temperature measurements at 17:00 h, participants completed a 5-min active warm-up before a ‘strength’ protocol. Participants performed three repetitions of right-hand grip strength, then three repetitions at each incremental load (40%, 60% and 80% of 1RM) for bench press and inclined leg press, with a 5-min recovery in between each repetition. A linear encoder was attached perpendicular to the movement, to the bar used for the exercises. Average power (AP), average force (AF), peak velocity (PV), distance (D) and time-to-peak velocity (tPV) were measured (MuscleLab software) during the concentric phase of the movements for each lift. Data were analysed using general linear models with repeated measures. The main findings were that SD reduced maximal grip (2.7%), bench press (11.2% AP, 3.3% AF and 9.4% PV) and leg press submaximal values (5.7% AP) with a trend for a reduction in AF (3.3% P = 0.06). Furthermore, RPE increased for measures of grip strength, leg and bench press during SD. Following a 1-h powernap (SD_N), values of grip and bench press improved to values similar in N, as did tiredness, alertness and sleepiness. There was a main effect for “load” on the bar for both bench and leg press where AP, AF, tPV values increased with load (P < 0.05) and PV decreased from the lightest to the heaviest load for both bench and leg press. An interaction of “load and condition” was present in leg press only, where the rate of change of AP is greater in the N than SD and SD_N conditions. In addition, for PV and tPV the rate of change was greater for SD_N than N or SD condition values. In summary, SD had a negative effect on grip strength and some components of bench and inclined leg press. The use of a 1-h power nap that ended 3 h before the “strength” assessment had a positive effect on weightlifting performance, subjective mood and ratings of tiredness.

Two-dimensional gel electrophoresis provides robust comparative analysis of skeletal muscle, but this technique is laborious and limited by its inability to resolve all proteins. In contrast, orthogonal separation by SDS-PAGE and reverse-phase liquid chromatography (RPLC) coupled to mass spectrometry (MS) affords deep mining of the muscle proteome, but differential analysis between samples is challenging due to the greater level of fractionation and the complexities of quantifying proteins based on the abundances of their tryptic peptides. Here we report simple, semi-automated and time efficient (i.e., 3 h per sample) proteome profiling of skeletal muscle by 1-dimensional RPLC electrospray ionisation tandem MS. Solei were analysed from rats (n = 5, in each group) bred as either high- or low-capacity runners (HCR and LCR, respectively) that exhibited a 6.4-fold difference (1,625 ± 112 m vs. 252 ± 43 m, p < 0.0001) in running capacity during a standardized treadmill test. Soluble muscle proteins were extracted, digested with trypsin and individual biological replicates (50 ng of tryptic peptides) subjected to LC-MS profiling. Proteins were identified by triplicate LC-MS/MS analysis of a pooled sample of each biological replicate. Differential expression profiling was performed on relative abundances (RA) of parent ions, which spanned three orders of magnitude. In total, 207 proteins were analysed, which encompassed almost all enzymes of the major metabolic pathways in skeletal muscle. The most abundant protein detected was type I myosin heavy chain (RA = 5,843 ± 897) and the least abundant protein detected was heat shock 70 kDa protein (RA = 2 ± 0.5). Sixteen proteins were significantly (p < 0.05) more abundant in HCR muscle and hierarchal clustering of the profiling data highlighted two protein subgroups, which encompassed proteins associated with either the respiratory chain or fatty acid oxidation. Heart-type fatty acid binding protein (FABPH) was 1.54-fold (p = 0.0064) more abundant in HCR than LCR soleus. This discovery was verified using selective reaction monitoring (SRM) of the y5 ion (551.21 m/z) of the doubly-charged peptide SLGVGFATR (454.19 m/z) of residues 23–31 of FABPH. SRM was conducted on technical replicates of each biological sample and exhibited a coefficient of variation of 20%. The abundance of FABPH measured by SRM was 2.84-fold greater (p = 0.0095) in HCR muscle. In addition, SRM of FABPH was performed in vastus lateralis samples of young and elderly humans with different habitual activity levels (collected during a previous study) finding FABPH abundance was 2.23-fold greater (p = 0.0396) in endurance-trained individuals regardless of differences in age. In summary, our findings in HCR/LCR rats provide protein-level confirmation for earlier transcriptome profiling work and show LC-MS is a viable means of profiling the abundance of almost all major metabolic enzymes of skeletal muscle in a highly parallel manner. Moreover, our approach is relatively more time efficient than techniques relying on orthogonal separations, and we demonstrate LC-MS profiling of the HCR/LCR selection model was able to highlight biomarkers that also exhibit differences in trained and untrained human muscle.

Purpose: The aim of this review was to assess current evidence regarding changes in cognitive function according to time-of-day (TOD) and assess the key components of research design related to manuscripts of chronobiological nature. Methods: An English-language literature search revealed 523 articles through primary database searches, and 1868 via organization searches/citation searching. The inclusion criteria were met by eleven articles which were included in the review. The inclusion criteria set were healthy adult males, a minimum of two timepoints including morning and evening, cognitive measures of performance, and peer-reviewed academic paper. Results: It was established that cognitive performance varies with TOD and the degree of difference is highly dependent on the type of cognitive task with differences ranging from 9.0 to 34.2% for reaction time, 7.3% for alertness, and 7.8 to 40.3% for attention. The type of cognitive function was a determining factor as to whether the performance was better in the morning, evening, or afternoon. Conclusion: Although some studies did not establish TOD differences, reaction time and levels of accuracy were highest in the evening. This implies that cognitive processes are complex, and existing research is contradictory. Some studies or cognitive variables did not show any measurable TOD effects, which may be due to differences in methodology, subjects involved, testing protocols, and confounding factors. No studies met all requirements related to chronobiological research, highlighting the issues around methodology. Therefore, future research must use a rigorous, approach, minimizing confounding factors that are specific to examinations of TOD.

Muscle force production and power output in active males, regardless of the site of measurement (hand, leg, or back), are higher in the evening than in the morning. This diurnal variation is attributed to motivational, peripheral and central factors, and higher core and, possibly, muscle temperatures in the evening. This study investigated whether increasing morning rectal temperatures to evening resting values, by active or passive warm-ups, leads to muscle force production and power output becoming equal to evening values in motivated subjects. Ten healthy active males (mean ± SD: age, 21.2 ± 1.9 yrs; body mass, 75.4 ± 8 kg; height, 1.76 ± .06 m) completed the study, which was approved by the University Ethics Committee. The subjects were familiarized with the techniques and protocol and then completed four sessions (separated by at least 48 h): control morning (07:30 h) and evening (17:30 h) sessions (with an active 5-min warm-up) and then two further sessions at 07:30 h but proceeded by an extended active or passive warm-up to raise rectal temperature to evening values. These last two sessions were counterbalanced in order of administration. During each trial, three measures of handgrip strength, isokinetic leg strength measurements (of knee flexion and extension at 1.05 and 4.19 rad.s⁻¹ through a 90° range of motion), and four measures of maximal voluntary contraction (MVC) on an isometric ergometer (utilizing the twitch-interpolation technique) were performed. Rectal and intra-aural temperatures, ratings of perceived exertion (RPE) and thermal comfort (TC) were measured. Measurements were made after the subjects had reclined for 30 min and after the warm-ups and prior to the measurement of handgrip and isokinetic and isometric ergometry. Muscle temperature was taken after the warm-up and immediately before the isokinetic and MVC measurements. Warm-ups were either active (cycle ergometer at 150 W) or passive (resting in a room at 35°C, relative humidity 45%). Data were analyzed using analysis of variance models with repeated measures. Rectal and intra-aural temperatures were higher at rest in the evening (.56°C and .74°C; p < .05) than in the morning, but there were no differences after the active or passive warm-ups, the subjects' ratings of thermal comfort reflecting this. Muscle temperatures also displayed significant diurnal variation, with higher values in the evening (∼.31°C; p < .05). Grip strength, isokinetic knee flexion for peak torque and peak power at 1.05 rad.s⁻¹, and knee extension for peak torque at 4.19 rad.s⁻¹ all showed higher values in the evening. All other measures of strength or power showed a trend to be higher in the evening ( .10 > p > .05). There was no significant effect of active or passive warm-ups on any strength or power variable, and subjects reported maximal values for effort for each strength measure. In summary, effects of time of day were seen in some measures of muscle performance but, in this population of motivated subjects, there was no evidence that increasing morning rectal temperature to evening values by active or passive warm-up increased muscle strength to evening values. (Author correspondence: B. J. Edwards@ljmu. ac. uk )

Numerous functional measures related to anaerobic performance display daily variation. The diversity of tests and protocols used to assess anaerobic performance related to diurnal effects and the lack of a standardized approach have hindered agreement in the literature. Therefore, the aim of the present study was to investigate and systematically review the evidence relating to time-of-day differences in anaerobic performance measures. The entire content of PubMed (MEDLINE), Scopus, SPORTDiscus® (via EBSCOhost) and Web of Science and multiple electronic libraries were searched. Only experimental research studies conducted in male adult participants aged ≥ 18 yrs before May 2021 were included. Studies assessing tests related to anaerobic capacity or anaerobic power between a minimum of two time-points during the day (morning vs evening) were deemed eligible. The primary search revealed that a total of 55 out of 145 articles were considered eligible and subsequently included. Thirty-nine studies assessed anaerobic power and twenty-five anaerobic capacity using different modes of exercise and test protocols. Forty-eight studies found several of their performance variables to display time-of-day effects, with higher values in the evening than the morning, while seven studies did not find any time-of-day significance in any variables which were assessed. The magnitude of difference is dependent on the modality and the exercise protocol used. Performance measures for anaerobic power found jump tests displayed 2.7 to 12.3% differences, force velocity tests ~8% differences, sprint tests 2.7 to 11.3% differences and 5-m multiple shuttle run tests 3.7 to 13.1% differences in favour of the evening. Performance measures for anaerobic capacity found Wingate test to display 1.8 to 11.7% differences and repeated sprint tests to display 3.4 to 10.2% differences. The only test not to display time-of-day differences was the running based anaerobic sprint test (RAST). Time-of-day variations in anaerobic performance has previously been partially explained by higher core-body and/or muscle temperature and better muscle contractile properties in the afternoon, although recent findings suggest that differences in methodology, motivation/arousal, habitual training times and chronotypes could provide additional explanations. There is a clear demand for a rigorous, standardised approach to be adopted by future investigations which control factors that specifically relate to investigations of time-of-day.