Live-cell three-dimensional single-molecule tracking reveals modulation of enhancer dynamics by NuRD.

To understand how the nucleosome remodeling and deacetylase (NuRD) complex regulates enhancers and enhancer-promoter interactions, we have developed an approach to segment and extract key biophysical parameters from live-cell three-dimensional single-molecule trajectories. Unexpectedly, this has revealed that NuRD binds to chromatin for minutes, decompacts chromatin structure and increases enhancer dynamics. We also uncovered a rare fast-diffusing state of enhancers and found that NuRD restricts the time spent in this state. Hi-C and Cut&Run experiments revealed that NuRD modulates enhancer-promoter interactions in active chromatin, allowing them to contact each other over longer distances. Furthermore, NuRD leads to a marked redistribution of CTCF and, in particular, cohesin. We propose that NuRD promotes a decondensed chromatin environment, where enhancers and promoters can contact each other over longer distances, and where the resetting of enhancer-promoter interactions brought about by the fast decondensed chromatin motions is reduced, leading to more stable, long-lived enhancer-promoter relationships.

What Can Pharmacological Models of Retinal Degeneration Tell Us?

Animal models with pharmacologically induced retinal degeneration including sodium iodate (NaIO3) and N-methyl-N-nitrosourea (MNU) have been extensively used in ophthalmic research to investigate retinal degeneration. NaIO3 induces degeneration of the retinal pigment epithelium (RPE) followed by photoreceptor (PRC) cell death, mimicking features of age-related macular degeneration. In contrast, MNU leads to rapid destruction of the PRCs only, enabling the use of the MNU model to investigate degeneration induced in retinitis pigmentosa. It has been shown that multiple cell death pathways are involved in the cell-specific effects of the toxins. Necrosis has been identified as the cause of the NaIO3-induced RPE loss. PRC degeneration in the described models is mainly induced by programmed cell death, indicated by the upregulation of conventional apoptosis initiator and effector caspases. However, recent research points to the additional involvement of caspase-independent processes as endoplasmic reticulum stress and calpain activation. Since there is still a substantial amount of contradictory hypotheses concerning triggers of cell death, the use of pharmacological models is controversial. Thereby, the advantages of such models like the application reaching across species and strains as well as modulation of onset and severity of damage are not exploited to a full extent. Thus, the present review aims to give more insight into the involved cell death pathways and discusses recent findings in the most widely used retinal degeneration models. It might facilitate further studies aiming to develop putative therapeutic approaches for retinal degenerative diseases including combinatory treatment with cell death inhibitors and cell transplantation therapy.

The Nucleosome Remodeling and Deacetylase Complex NuRD Is Built from Preformed Catalytically Active Sub-modules.

The nucleosome remodeling deacetylase (NuRD) complex is a highly conserved regulator of chromatin structure and transcription. Structural studies have shed light on this and other chromatin modifying machines, but much less is known about how they assemble and whether stable and functional sub-modules exist that retain enzymatic activity. Purification of the endogenous Drosophila NuRD complex shows that it consists of a stable core of subunits, while others, in particular the chromatin remodeler CHD4, associate transiently. To dissect the assembly and activity of NuRD, we systematically produced all possible combinations of different components using the MultiBac system, and determined their activity and biophysical properties. We carried out single-molecule imaging of CHD4 in live mouse embryonic stem cells, in the presence and absence of one of core components (MBD3), to show how the core deacetylase and chromatin-remodeling sub-modules associate in vivo. Our experiments suggest a pathway for the assembly of NuRD via preformed and active sub-modules. These retain enzymatic activity and are present in both the nucleus and the cytosol, an outcome with important implications for understanding NuRD function.

[Stem cell therapy for retinal diseases]. / Stammzelltherapie für Netzhauterkrankungen.

BACKGROUND: Due to an ageing population the incidence and prevalence of retinal diseases and visual disabilities will continue to grow. A great number of patients would principally be able to benefit from a stem cell-based therapy. OBJECTIVES: To introduce readers to the terminology and current concepts associated with stem cell therapy in ocular research and to provide an overview of the current status of preclinical and clinical research. MATERIAL AND METHODS: We performed a systematic review of relevant entries on ocular stem cell therapy for retinal diseases in PubMed and ClinicalTrials.gov. Differences between various stem cell types are displayed systematically, followed by a discussion of preclinical studies. Translational aspects are highlighted leading to the first clinical trials, including surgical and ethical facets. RESULTS: In preclinical studies, photoreceptor cell precursors and retinal pigment epithelium (RPE) cells were differentiated and subretinally transplanted into animal models. Besides exclusion of a teratoma formation, some functional improvements were also observed. Intraocular transplantation of stem cell-derived RPE cells was the first successful clinical application of pluripotent stem cells in man. CONCLUSION: Promising results of preclinical and clinical studies have identified important challenges and confirmed the potential of stem cell therapy for ophthalmology.

Combined aerobic and resistance training in breast cancer survivors: A randomized, controlled pilot trial.

The purpose of this pilot study was to examine the effects of a combined cardiorespiratory and resistance exercise training program of short duration on the cardiorespiratory fitness, strength endurance, task specific functional muscle capacity, body composition and quality of life (QOL) in women breast cancer survivors. Sixteen subjects were randomly assigned to either a training (n = 8; age: 50 +/- 5 yrs) or control non-exercising group (n = 8; age: 51 +/- 10 yrs). The training group followed an 8-week exercise program consisting of 3 weekly sessions of 90-min duration, supervised by an experienced investigator and divided into resistance exercises and aerobic training. Before and after the intervention period, all of the subjects performed a cardiorespiratory test to measure peak oxygen uptake (VO2peak), a dynamic strength endurance test (maximum number of repetitions for chest and leg press exercise at 30 - 35 % and 100 - 110 % of body mass, respectively) and a sit-stand test. Quality of life was assessed using the European Organization for Research and Treatment of Cancer QLQ-C30 (EORTC-C30) questionnaire. In response to training, QOL, VO2peak (mean 3.9 ml/kg/min; 95 % CI, 0.93, 6.90) performance in leg press (17.9 kg; 95 % CI, 12.8, 22.4) and sit-stand test (- 0.67 s; 95 % CI, - 0.52, - 1.2) improved (p < or = 0.05). We observed no significant changes in the control group. Combined cardiorespiratory and resistance training, even of very brief duration, improves the QOL and the overall physical fitness of women breast cancer survivors.

A robust characterization of retinoic acid response elements based on a comparison of sites in three species.

The availability of high-throughput genomic sequencing has allowed us to construct a more robust characterization of retinoic acid response elements than was possible in the past. We located human, mouse, and rat homologs for each of 51 well-documented, conserved retinoic acid response elements. Mathematical and statistical analyses of these 153 sites, 78 of which are new, shows that 92% of response elements have direct-repeat symmetry, but that only 76% exhibit canonical spacing attributes. While the familiar '(a/g)g(g/t)tca' hexamer motif is upheld, the more relaxed sequence, '(a/g)g(g/t)(g/t)(g/c)a', represents a 10% consensus. Sites are as likely to be on the coding strand as on the non-coding strand, and 86% of them are in upstream locations. From a statistical point of view, DR1 elements are fundamentally different from DR2 and DR5 elements, but this is only evident in the 5' hexamer. While there is considerable variation in core positions, and while no nucleotide can be considered forbidden at any position, variation among species at a fixed locus appears surprisingly constrained once a functional site has been attained.

Effects of the rotor pedalling system on the performance of trained cyclists during incremental and constant-load cycle-ergometer tests.

The aim of this study was to determine the effects of Rotor, a new cycle crank configuration that effectively allows the pedals to move independently throughout the duty cycle, on indices of endurance cycling performance in trained cyclists. Ten cyclists (5 Rotor users and 5 non-users; age (mean +/- SD): 22 +/- 5 y; VO(2)max: 69.5 +/- 5.1 mL. kg(-1).min(-1)) volunteered to participate in the study. On four separate days, the subjects performed four cycle-ergometer tests, i.e. two incremental tests and two 20-min tests. An imposed crank rate of 75 rev.min(-1) was used during all tests. The incremental protocol started at 112.5 W, and the power output was increased by 37.5 W every 3 min until volitional exhaustion. The 20-min tests were performed at a fixed power output equivalent to 80 % of the highest power output that the cyclists maintained for a complete 3-min period during incremental tests. Both types of tests were performed with the conventional crank system and the Rotor following a counter-balanced, cross-over design. Gas exchange parameters were measured in all the tests and blood lactate was determined at the end of each 3-min period (incremental tests) and at the end of the 20-min tests. A three factor (pedalling system used during the tests x habitual pedalling system x power output [incremental tests] or time [20-min tests]) ANOVA with repeated measures on power output (incremental tests) or time (20-min tests) was used to analyse several indices of performance, e.g. peak power output, VO(2)max, lactate threshold, onset of blood lactate accumulation, economy, delta, and gross efficiency. No differences (p > 0.05) were found between the Rotor and conventional systems for any of the aforementioned variables. It seems that the theoretical advantage brought about by the Rotor system, i.e. improved contra-lateral cooperation of both legs, would be minimized in trained cyclists. Although field studies are needed to assess the possible implications, in terms of actual racing, of the new system, commonly used indicators of endurance cycling performance do not seem to be improved with the Rotor in trained cyclists.

Reliability of a combined 3-min constant load and performance cycling test.

Most fitness assessments either use a constant load to exhaustion (exercise capacity test) or an "all-out" effort (performance test). The purpose of this study was to determine the reliability of a high-intensity assessment that combined a constant load element with a performance test. Ten moderately trained male cyclists completed a ramp test to voluntary exhaustion in order to measure maximum minute power output (mean +/- s, 349.3 w +/- 55.0 w). On two other occasions subjects cycled at a constant load at maximum minute power output for 2-min immediately followed by a 1-min performance test. All tests were conducted on the subjects' own bicycles using a Kingcycle trade mark test rig. Power output was measured each second using SRM trade mark Power Cranks. The data were analysed by measuring the reliability of each 30 s of the 3-min test together with the peak power and the peak cadence achieved in the performance element of the test. There was no systematic bias in the data from trial 1 to trial 2 for any of the 6, 30 s blocks of the test, the peak power (mean, 95 % CI, 413.8 w, 357.8 - 469.7 w and 403.8 w, 339.9 - 467.6 w, trial 1 and trial 2, respectively) or peak cadence (95.0 rev x min(-1), 89.5 - 100.5 rev x min(-1) and 95.1 rev x min(-1), 90.0 - 100.1 rev x min(-1), trial 1 and trial 2, respectively). Mean (+/- s) total distance over the 3-min was 2.23 +/- 0.23 km and 2.26 +/- 0.26 km for trial 1 and trial 2 respectively (p > 0.05). The coefficients of variation ranged from 0.9 - 5.4 % and the intraclass correlation coefficients ranged from 0.96 - 0.99. It is concluded that in moderately trained subjects, the 3-min combination test provides reliable data and could therefore be used for short-term, high-intensity cycling intervention studies.

Heart rate responses of women aged 23-67 years during competitive orienteering.

OBJECTIVES: To compare the heart rate responses of women orienteers of different standards and to assess any relation between heart rate responses and age. METHODS: Eighteen competitive women orienteers completed the study. They were divided into two groups: eight national standard orienteers (ages 23-67 years); 10 club standard orienteers (ages 24-67 years). Each participant had her heart rate monitored during a race recognised by the British Orienteering Federation. Peak heart rate (HR(PEAK)), mean heart rate (HR(MEAN)), standard deviation of her heart rate during each orienteering race (HR(SD)), and mean change in heart rate at each control point (DeltaHR(CONTROL)) were identified. The data were analysed using analysis of covariance with age as a covariate. RESULTS: National standard orienteers displayed a lower within orienteering race standard deviation in heart rate (6 (2) v 12 (2) beats/min, p<0.001) and a lower DeltaHR(CONTROL) (5 (1) v 17 (4) beats/min, p<0.001). The mean heart rate during competition was higher in the national standard group (170 (11) v 158 (11) beats/min, p = 0.025). The HR(MEAN) for the national and club standard groups were 99 (8)% and 88 (9)% of their age predicted maximum heart rate (220-age) respectively. All orienteers aged >55 years (n = 4) recorded HR(MEAN) greater than their age predicted maximum. CONCLUSIONS: The heart rate responses indicate that national and club standard women orienteers of all ages participate in the sport at a vigorous intensity. The higher DeltaHR(CONTROL) of club standard orienteers is probably due to failing to plan ahead before arriving at the controls and this, coupled with slowing down to navigate or relocate when lost, produced a higher HR(SD).

Heart rate responses of male orienteers aged 21-67 years during competition.

Orienteering is a sport in which it is common for most participants to be aged over 40 years, but research into the demands of the sport has focused almost exclusively on elite participants aged 21-35 years. The aim of the present study was to examine the heart rate responses of older male orienteers. Thirty-nine competitive male orienteers were divided into three groups: group 1 (international competitive standard, n = 11, age 21-67 years), group 2 (national competitive standard, n = 15, age 24-66 years) and group 3 (club competitive standard, n = 13, age 23-60 years). Each participant had his heart rate monitored during two orienteering races of contrasting technical difficulty. The results were analysed using analysis of covariance, with age as a covariate, and Pearson product-moment correlation coefficients to determine whether age was related to the observed heart rate responses. The groups did not differ in their peak (175 +/- 12 beats x min(-1), P = 0.643) or mean (159 +/- 13 beats x min(-1), P = 0.171) heart rates during the races. There was a decline of 6 beats x min(-1) x decade(-1) (P = 0.001) for peak heart rate and 5 beats x min(-1) x decade(-1) (P < 0.001) for mean heart rate. Mean heart rates were 86 +/- 6% of the participants' maximal heart rates and were not associated with age. The orienteers in group 1 displayed a lower (P < 0.005) within-race standard deviation in heart rate (6 +/- 2 beats x min(-1)) than those in groups 2 and 3 (10 +/- 3 and 10 +/- 4 beats x min(-1), respectively). In conclusion, the mean heart rates indicated that all three groups of orienteers ran at a relative high intensity and the international competitive standard orienteers displayed a less variable heart rate, which may have been related to fewer instances of slowing down to relocate and being able to navigate while running at relatively high speeds.

Reliability of mean power recorded during indoor and outdoor self-paced 40 km cycling time-trials.

The purpose of this study was to assess reliability of both indoor and outdoor 40 km time-trial cycling performance. Eight trained cyclists completed three indoor 40 km time-trials on an air-braked ergometer (Kingcycle) and three outdoor 40 km time-trials on a local course. Power output was measured for all trials using the SRM powermeter. Mean performance time across three indoor trials was 54.21 +/- 2.59 (min:sec) and was significantly different (P<0.05) to mean time across three outdoor trials (57.29 +/- 3.22 min:sec). However, there was no significant difference (P = 0.34) for mean power across three indoor trials (303+/-35W) when compared to outdoor performances (312 +/- 23 W). Within-subject variation for mean power output expressed as a coefficient of variation (CV) improved in both indoors and outdoors for trials 2 and 3 (CV = 1.9%, 95% CI 1.0 - 3.4 and CV = 2.1 %, 95 % CI 1.1 - 3.8) when compared to trials 1 and 2 (CV=2.1%, 95% CI 1.2-3.8 and CV=2.4%, 95% CI 1.3-4.3). These findings indicate that power output measured using the SRM powermeter is highly reproducible for both laboratory-based and actual 40 km time-trial cycling performance.

Differences between the sexes and age-related changes in orienteering speed.

The aim of this study was to assess the effects of the age and sex of the competitor on orienteering speed during competitive events. The results of the fastest three male and fastest three female competitors in each 5-year age band (21-79 years), from four national orienteering events, were analysed. The data for age and orienteering speed were log-transformed and regression analyses were conducted to determine the relationships between age and sex and orienteering speed. For comparison, data for the fastest Great Britain finisher in the 10,000-m track and 10-km cross-country events for age groups 40-69 years at the World Masters Championships were also analysed. The results showed that, before the age of 40 years, there was no substantial slowing in orienteering speed for males (0.5-4.2% per decade) but a moderate decline (4.7-10.0% per decade) for females. After the age of 45 years, the orienteering speed of males and females slowed by 13+/-2% and 16+/-4% per decade (mean +/- s), respectively, until around the age of 69, after which the deterioration was accentuated. The orienteering speed of the females was 81+/-4, 74+/-6 and 69+/-7% that of the males at ages 21, 45 and 65 years, respectively. The magnitudes of the age-related slowing of orienteering speed and of the difference in orienteering speed between males and females aged 45 years and over were greater than those reported for the other endurance running events. This may reflect the physical demands of running in orienteering terrain, tactical and cognitive aspects of the sport, or sociocultural aspects of the participating population.

Reliability of an air-braked ergometer to record peak power during a maximal cycling test.

PURPOSE: To assess the reliability of the Kingcycle ergometer, this study compared peak power recorded using a Kingcycle and SRMTM power meters during Kingcycle maximal aerobic power tests. METHODS: The study was completed in two parts: for part 1, nine subjects completed three maximal tests with a stabilizing kit attached to the Kingcycle rig and calibration of the Kingcycle checked against SRM (MAP(C)); and for part 2, nine subjects completed two maximal tests without the stabilizing kit and the Kingcycle calibrated using the standard procedure (MAP(S)). Each MAP(C) test was separated by 1 wk; however, MAPs tests were separated by 54 +/- 32 d, (mean +/- SD). Testing procedure was repeated for each MAP and peak power output was calculated as the highest average power output recorded during any 60-s period of the MAP test using the Kingcycle (King(PPO)) and SRM (SRM(PPO)). RESULTS: Coefficient of variations (CVs) for King(PPO) were larger than those of SRM(PPO); 2.0% (95%CI = 1.5-3.0) versus 1.3% (95%CI = 1.0-2.0) and 4.6% (95%CI = 2.7-7.6) versus 3.6% (95%CI = 2.1-6.0) for MAPC( and MAP(S), respectively. During all tests, King(PPO) was higher than SRM(PPO) by an average of approximately 10% (P < 0.001). CONCLUSIONS: Investigators should be aware of the discrepancy between the two systems when assessing peak power and that SRM cranks provide a more reproducible measure of peak power than the Kingcycle ergometer.

Peak power predicts performance power during an outdoor 16.1-km cycling time trial.

PURPOSE: To assess i) the reproducibility of peak power output recorded during a maximal aerobic power test (MAP), and ii) its validity to predict endurance performance during a field based 16.1-km time trial (16.1-km TT). METHODS: Two studies were completed: for part I, nine subjects performed three MAP tests; for part II, 16 subjects completed a MAP test and 16.1-km TT. Power output was recorded using an SRM power meter and was calculated as peak power output (PPO) recorded during 60 s of MAP and mean power output for the 16.1-km TT (16.1-km TT(PO)). RESULTS: There was no difference between PPO recorded during the three MAP trials, mean coefficient of variation for individual cyclists was 1.32% (95%CI = 0.97-2.03), and test-retest reliability expressed as an intraclass correlation coefficient was 0.99 (95%CI = 0.96-1.00). A highly significant relationship was found between PPO and 16.1-km TT(PO) (r = 0.99, P < 0.001) but not for PPO and 16.1-km TT time (r = 0.46. P > 0.05). CONCLUSION: The results show that PPO affords a valid and reliable measure of endurance performance which can be used to predict average power during a 16.1-km TT but not performance time.

The validity of power output recorded during exercise performance tests using a Kingcycle air-braked cycle ergometer when compared with an SRM powermeter.

This study assessed the validity of power output recorded using an air-braked cycle ergometer (Kingcycle) when compared with a power measuring crankset (SRM). For part one of the study thirteen physically active subjects completed a continuous incremental exercise test (OBLA), for part two of the study twelve trained cyclists completed two tests; a maximal aerobic power test (MAP) and a 16.1 km time-trial (16.1 km TT). The following were compared; the peak power output (PPO) recorded for 1 min during MAP, the average power output for the duration of the time-trial and power output recorded during each stage of OBLA. For all tests, power output recorded using Kingcycle was significantly higher than SRM (P < 0.001). Ratio limits of agreement between SRM and Kingcycle for OBLA showed a bias (P < 0.00) of 0.90 (95%CI = 0.90-0.91) with a random error of X or / 1.07, and for PPO and 16.1 km TT ratio limits of agreement were 0.90 (95%CI = 0.88-0.92) X or / 1.07 and 0.92 (95% CI = 0.90-0.94) X or / 1.07, respectively. These data revealed that the Kingcycle ergometry system did not provide a valid measure of power output when compared with SRM.

Assessment of blood lactate: practical evaluation of the Biosen 5030 lactate analyzer.

PURPOSE: The aim of this study was to assess the validity and reliability of the Biosen 5030 lactate analyzer compared with a YSI 2300 lactate analyzer and a Kodak Ektachem DTII in a practical laboratory study context. METHODS: To assess validity, 144 triplicate capillarized blood samples, across a range of values, were analyzed using the three analyzers. To assess reliability a further 665 samples were repeat analyzed. Temporal stability was determined by the reanalysis of resting and maximal exercise blood samples, after a period of storage ranging from 7 to 20 h, at room temperature. To measure inter- and intra-investigator reliability, 20 resting samples were taken from three different subjects by different investigators and a coefficient of variation was determined. RESULTS: There were strong relationships between the Biosen, the YSI (r2 = 0.97), and the Kodak Ektachem (r2 = 0.91). An analysis of Biosen compared with YSI revealed a positive bias of 0.37 mmol x L(-1) (95% limits of agreement, -0.85 to 1.59 mmol x L(-1)). The test-retest reliability correlation was significant (r2 = 0.99, P < 0.05), but a paired t-test revealed a small (0.03 mmol x L(-1), P < 0.05) significant difference. The coefficient of variation from the three investigators across the 20 samples ranged from 1.3 to 3%. Blood lactate concentration in resting blood samples did significantly increase in value (0.2 mmol x L(-1), P < 0.05) after 7-h exposure to the air, whereas there was no change in maximal exercise blood lactate values after 20-h exposure to the air. CONCLUSIONS: In a practical context, the Biosen 5030 lactate analyzer was comparable to the other analyzers giving fast reliable measures of blood lactate concentrations over the full range of values, which remained stable over extended periods at room temperature.

Talbot array illuminator for single-shot measurements of laser-induced-damage thresholds of thin-film coatings.

A highly efficient Talbot array illuminator for single-shot, laser-induced-damage test measurements of optical thin-film coatings is proposed. With a periodic binary phase grating, a laser beam is transformed into an ensemble of Gaussian-like spots, which are known as the Fresnel image of the grating. For this purpose hexagonal phase gratings were fabricated and analyzed. With a peak fluence distribution of approximately 1 order of magnitude, the damage threshold of thin films can be deduced by use of the data from only a single shot.

Experimental determination of the fundamental-mode diameter in solid-state lasers.

A method to determine directly the radius of the fundamental mode in a laser crystal has been developed. The radius is measured by comparison of the distribution of the spontaneous emission in the pumped region during laser operation with the fluorescence distribution without laser emission. Measuring the mode radius with various pump powers enables one to optimize the overlap between the pump and the cavity beam and to determine the dioptric power of the thermally induced lens.

Carbohydrate ingestion improves endurance performance during a 1 h simulated cycling time trial.

This study examined the effect of carbohydrate ingestion on metabolic and performance-related responses during and after a simulated 1 h cycling time trial. Eight trained male cyclists (VO2 peak = 66.5 ml kg-1 min-1) rode their own bicycles mounted on a windload simulator to imitate real riding conditions. At a self-selected maximal pace, the cyclists performed two 1 h rides (separated by 7 days) and were fed either an 8% carbohydrate or placebo solution. The beverages were administered 25 min before (4.5 ml kg-1) and at the end (4.5 ml kg-1) of the ride. With carbohydrate feeding, plasma glucose tended (P = 0.21) to rise before the time trial. Compared with rest, the plasma glucose concentration decreased significantly (P < 0.05) at the end of both rides, with no statistically significant difference being observed between treatments. Thereafter, plasma glucose increased significantly (P < 0.05) at 15 and 30 min into recovery and was significantly higher at 30 min during the carbohydrate trial compared with the placebo trial. No significant changes in plasma free fatty acids were observed during the ride. However, a significant increase (P < 0.05) in free fatty acids was found at 15 and 30 min into recovery, with no difference between trials. Mean power output was significantly (P < 0.05) greater during the carbohydrate compared with the placebo trial (mean +/- S.E.: 277 +/- 3 and 269 +/- 3 W, respectively). The greater distance covered in the carbohydrate compared with the placebo trial (41.5 +/- 1.06 and 41.0 +/- 1.06 km, respectively; P < 0.05) was equivalent to a 44 s improvement. We conclude that pre-exercise carbohydrate ingestion significantly increases endurance performance in trained cyclists during a 1 h simulated time trial. Although the mechanism for this enhancement in performance with carbohydrate ingestion cannot be surmised from the present results, it could be related to a higher rate of carbohydrate oxidation, or to favourable effects of carbohydrate ingestion on the central component of fatigue.