Jonathan Fulford

Dietary nitrate supplementation reduces the O2 cost of walking and running: a placebo-controlled study

Dietary supplementation with beetroot juice (BR) has been shown to reduce resting blood pressure and the O(2) cost of submaximal exercise and to increase tolerance to high-intensity cycling. We tested the hypothesis that the physiological effects of BR were consequent to its high NO(3)(-) content per se, and not the presence of other potentially bioactive compounds. We investigated changes in blood pressure, mitochondrial oxidative capacity (Q(max)), and physiological responses to walking and moderate- and severe-intensity running following dietary supplementation with BR and NO(3)(-)-depleted BR [placebo (PL)]. After control (nonsupplemented) tests, nine healthy, physically active male subjects were assigned in a randomized, double-blind, crossover design to receive BR (0.5 l/day, containing ∼6.2 mmol of NO(3)(-)) and PL (0.5 l/day, containing ∼0.003 mmol of NO(3)(-)) for 6 days. Subjects completed treadmill exercise tests on days 4 and 5 and knee-extension exercise tests for estimation of Q(max) (using (31)P-magnetic resonance spectroscopy) on day 6 of the supplementation periods. Relative to PL, BR elevated plasma NO(2)(-) concentration (183 ± 119 vs. 373 ± 211 nM, P < 0.05) and reduced systolic blood pressure (129 ± 9 vs. 124 ± 10 mmHg, P < 0.01). Q(max) was not different between PL and BR (0.93 ± 0.05 and 1.05 ± 0.22 mM/s, respectively). The O(2) cost of walking (0.87 ± 0.12 and 0.70 ± 0.10 l/min in PL and BR, respectively, P < 0.01), moderate-intensity running (2.26 ± 0.27 and 2.10 ± 0.28 l/min in PL and BR, respectively, P < 0.01), and severe-intensity running (end-exercise O(2) uptake = 3.77 ± 0.57 and 3.50 ± 0.62 l/min in PL and BL, respectively, P < 0.01) was reduced by BR, and time to exhaustion during severe-intensity running was increased by 15% (7.6 ± 1.5 and 8.7 ± 1.8 min in PL and BR, respectively, P < 0.01). In contrast, relative to control, PL supplementation did not alter plasma NO(2)(-) concentration, blood pressure, or the physiological responses to exercise. These results indicate that the positive effects of 6 days of BR supplementation on the physiological responses to exercise can be ascribed to the high NO(3)(-) content per se.

Dietary nitrate reduces muscle metabolic perturbation and improves exercise tolerance in hypoxia

Non‐Technical Summary  Reduced atmospheric O2 availability (hypoxia) impairs muscle oxidative energy production and exercise tolerance. We show that dietary supplementation with inorganic nitrate reduces markers of muscle fatigue and improves high‐intensity exercise tolerance in healthy adults inhaling air containing 14.5% O2. In the body, nitrate can be converted to nitrite and nitric oxide. These molecules can improve muscle efficiency and also dilate blood vessels allowing more O2 to be delivered to active muscle. These results suggest that dietary nitrate could be beneficial during exercise at moderate to high altitude and in conditions where O2 delivery to muscle is reduced such as in pulmonary, cardiovascular and sleep disorders.

Influence of hyperoxia on muscle metabolic responses and the power–duration relationship during severe‐intensity exercise in humans: a 31P magnetic resonance spectroscopy study

Severe‐intensity constant‐work‐rate exercise results in the attainment of maximal oxygen uptake, but the muscle metabolic milieu at the limit of tolerance (Tlim) for such exercise remains to be elucidated. We hypothesized that Tlim during severe‐intensity exercise would be associated with the attainment of consistently low values of intramuscular phosphocreatine ([PCr]) and pH, as determined using 31P magnetic resonance spectroscopy, irrespective of the work rate and the inspired O2 fraction. We also hypothesized that hyperoxia would increase the asymptote of the hyperbolic power–duration relationship (the critical power, CP) without altering the curvature constant (W′). Seven subjects (mean ±s.d., age 30 ± 9 years) completed four constant‐work‐rate knee‐extension exercise bouts to the limit of tolerance (range, 3–10 min) both in normoxia (N) and in hyperoxia (H; 70% O2) inside the bore of 1.5 T superconducting magnet. The [PCr] (∼5–10% of resting baseline) and pH (∼6.65) at the limit of tolerance during each of the four trials was not significantly different either in normoxia or in hyperoxia. At the same fixed work rate, the overall rate at which [PCr] fell with time was attenuated in hyperoxia (mean response time: N, 59 ± 20 versus H, 116 ± 46 s; P < 0.05). The CP was higher (N, 16.1 ± 2.6 versus H, 18.0 ± 2.3 W; P < 0.05) and the W′ was lower (N, 1.92 ± 0.70 versus H, 1.48 ± 0.31 kJ; P < 0.05) in hyperoxia compared with normoxia. These data indicate that Tlim during severe‐intensity exercise is associated with the attainment of consistently low values of muscle [PCr] and pH. The CP and W′ parameters of the power–duration relationship were both sensitive to the inspiration of hyperoxic gas.

Effects of short-term dietary nitrate supplementation on blood pressure, O2 uptake kinetics, and muscle and cognitive function in older adults

Dietary nitrate (NO(3)(-)) supplementation has been shown to reduce resting blood pressure and alter the physiological response to exercise in young adults. We investigated whether these effects might also be evident in older adults. In a double-blind, randomized, crossover study, 12 healthy, older (60-70 yr) adults supplemented their diet for 3 days with either nitrate-rich concentrated beetroot juice (BR; 2 × 70 ml/day, ∼9.6 mmol/day NO(3)(-)) or a nitrate-depleted beetroot juice placebo (PL; 2 × 70 ml/day, ∼0.01 mmol/day NO(3)(-)). Before and after the intervention periods, resting blood pressure and plasma [nitrite] were measured, and subjects completed a battery of physiological and cognitive tests. Nitrate supplementation significantly increased plasma [nitrite] and reduced resting systolic (BR: 115 ± 9 vs. PL: 120 ± 6 mmHg; P < 0.05) and diastolic (BR: 70 ± 5 vs. PL: 73 ± 5 mmHg; P < 0.05) blood pressure. Nitrate supplementation resulted in a speeding of the Vo(2) mean response time (BR: 25 ± 7 vs. PL: 28 ± 7 s; P < 0.05) in the transition from standing rest to treadmill walking, although in contrast to our hypothesis, the O(2) cost of exercise remained unchanged. Functional capacity (6-min walk test), the muscle metabolic response to low-intensity exercise, brain metabolite concentrations, and cognitive function were also not altered. Dietary nitrate supplementation reduced resting blood pressure and improved Vo(2) kinetics during treadmill walking in healthy older adults but did not improve walking or cognitive performance. These results may have implications for the enhancement of cardiovascular health in older age.

Fetal Brain Activity in Response to a Visual Stimulus

Previous studies have demonstrated the use of functional magnetic resonance imaging (fMRI) to assess fetal brain activity. To extend these studies, a fetal fMRI experiment using a visual stimulus has been performed at 0.5 T. This used a block fMRI paradigm with a bright, constant‐intensity light source being shone at the maternal abdomen for 8 sec followed by 16 sec of darkness. This was repeated typically 40 times on nine subjects all of whom were greater than 36 weeks gestational age. Of these, one could not be analysed due to motion, three did not show significant activation, and five showed significant activation (P < 0.0085). In all cases, activation was localised within the frontal cortex. Exact localisation was difficult but this may correspond to the frontal eye fields and dorsolateral prefontal cortex. In no cases was significant activation present within the occipital region as would have been expected and was observed in 2/8 adult subjects. Hum. Brain Mapping 20:239–245, 2003.

An in-frame deletion at the polymerase active site of POLD1 causes a multisystem disorder with lipodystrophy

DNA polymerase δ, whose catalytic subunit is encoded by POLD1, is responsible for lagging-strand DNA synthesis during DNA replication. It carries out this synthesis with high fidelity owing to its intrinsic 3′- to 5′-exonuclease activity, which confers proofreading ability. Missense mutations affecting the exonuclease domain of POLD1 have recently been shown to predispose to colorectal and endometrial cancers. Here we report a recurring heterozygous single-codon deletion in POLD1 affecting the polymerase active site that abolishes DNA polymerase activity but only mildly impairs 3′- to 5′-exonuclease activity. This mutation causes a distinct multisystem disorder that includes subcutaneous lipodystrophy, deafness, mandibular hypoplasia and hypogonadism in males. This discovery suggests that perturbing the function of the ubiquitously expressed POLD1 polymerase has unexpectedly tissue-specific effects in humans and argues for an important role for POLD1 function in adipose tissue homeostasis.

Antenatal determination of fetal brain activity in response to an acoustic stimulus using functional magnetic resonance imaging.

Functional magnetic resonance imaging (fMRI) is now a well‐established technique for directly identifying adult brain activity. This study builds on earlier pilot work that showed that fMRI could provide direct evidence of fetal brain cortical activation in response to an auditory stimulus. The new work presented here aims to assess the sensitivity of this technique in a larger sample group. This article includes a specific discussion of the methodology required for fetal fMRI. Sixteen pregnant subjects were scanned between 37 and 41 weeks gestation, 12 had an auditory stimulus applied to the maternal abdomen (study group) and 4 had an auditory stimulus applied to the mothers ears (control group). Two of twelve (2/12) study‐group patients experienced back pain so that the experiment was abandoned; 4/12 showed significant activation (P < 0.005) in one or both of the temporal lobes; 1/12 showed significant activation in the frontal lobe. A susceptibility artifact at the interface between the maternal bowel and the fetus affected 3/12 data sets. No significant activation was found in 3/4 of the control cases, and 1/4 could not be analyzed due to a susceptibility artifact. Hum. Brain Mapping 12:94–99, 2001.

IN VITRO AND PRELIMINARY IN VIVO VALIDATION OF ECHO PARTICLE IMAGE VELOCIMETRY IN CAROTID VASCULAR IMAGING

Noninvasive, easy-to-use and accurate measurements of wall shear stress (WSS) in human blood vessels have always been challenging in clinical applications. Echo particle image velocimetry (Echo PIV) has shown promise for clinical measurements of local hemodynamics and wall shear rate. Thus far, however, the method has only been validated under simple flow conditions. In this study, we validated Echo PIV under in vitro and in vivo conditions. For in vitro validation, we used an anatomically correct, compliant carotid bifurcation flow phantom with pulsatile flow conditions, using optical particle image velocimetry (optical PIV) as the reference standard. For in vivo validation, we compared Echo PIV-derived 2-D velocity fields obtained at the carotid bifurcation in five normal subjects against phase-contrast magnetic resonance imaging (PC-MRI)-derived velocity measurements obtained at the same locations. For both studies, time-dependent, 2-D, two-component velocity vectors; peak/centerline velocity, flow rate and wall shear rate (WSR) waveforms at the common carotid artery (CCA), carotid bifurcation and distal internal carotid artery (ICA) were examined. Linear regression, correlation analysis and Bland-Altman analysis were used to quantify the agreement of different waveforms measured by the two techniques. In vitro results showed that Echo PIV produced good images of time-dependent velocity vector maps over the cardiac cycle with excellent temporal (up to 0.7 ms) and spatial (∼0.5 mm) resolutions and quality, comparable with optical PIV results. Further, good agreement was found between Echo PIV and optical PIV results for velocity and WSR measurements. In vivo results also showed good agreement between Echo PIV velocities and phase contrast MRI velocities. We conclude that Echo PIV provides accurate velocity vector and WSR measurements in the carotid bifurcation and has significant potential as a clinical tool for cardiovascular hemodynamics evaluation.

Fetal brain activity and hemodynamic response to a vibroacoustic stimulus

Previous studies have demonstrated the practicality of using functional magnetic resonance imaging (fMRI) techniques to assess fetal brain activity. The purpose of this study was to compare the fetal hemodynamic response to that of the adult. Seventeen pregnant subjects, all of whom were at more than 36 weeks gestation were scanned while the fetus was exposed to a vibroacoustic stimulus. Thirteen adult subjects were scanned with an equivalent acoustic stimulus. Of the fetal subjects, two could not be analyzed due to technical problems, eight did not show significant activation, and seven showed significant activation. In all cases, activation was localized within the temporal region. Measures of fetal hemodynamic responses revealed an average time to peak (ttp) of 7.36 ± 0.94 sec and an average percentage change of 2.67 ± 0.93%. In contrast, activation was detected in 5 of 13 adults with an average ttp of 6.54 ± 0.54 sec and an average percentage change of 1.02 ± 0.40%. The measurement of changes in the fetal hemodynamic response may be important in assessing compromised pregnancies. Hum. Brain Mapping 22:118–123, 2004.

Muscle metabolic determinants of exercise tolerance following exhaustion: relationship to the “critical power”

We tested the hypothesis that muscle high-energy phosphate compounds and metabolites related to the fatigue process would be recovered after exhaustion during recovery exercise performed below but not above critical power (CP) and that these changes would influence the capacity to continue exercise. Eight male subjects completed single-leg, knee-extension exercise to exhaustion (for ∼180 s) on three occasions, followed by a work-rate reduction to severe-intensity exercise, heavy-intensity exercise (<CP), or a 10-min passive recovery period, in random order. The muscle metabolic responses to exercise were assessed using (31)P magnetic resonance spectroscopy. There was a significant difference between the sustainable exercise duration during the recovery from exhaustive exercise between the <CP and >CP conditions (at least 10 min and 39 ± 31 s, respectively; P < 0.05). During passive recovery and <CP recovery exercise, muscle phosphocreatine concentration ([PCr]) increased rapidly after the exhaustion point, reaching ∼96% and ∼76% of baseline values, respectively, after 10 min (P < 0.05). Moreover, pH increased abruptly, reaching 7.0 ± 0.0 and 7.0 ± 0.2, respectively, after 10 min recovery (P < 0.05). However, during >CP recovery exercise, neither muscle [PCr] nor pH recovered, reaching ∼37% of the initial baseline and 6.6 ± 0.2, respectively. These results indicate that the muscle metabolic dynamics in recovery from exhaustive >CP differ according to whether the recovery exercise is performed below or above the CP. These findings confirm the importance of the CP as an intramuscular metabolic threshold that dictates the accumulation of fatigue-related metabolites and the capacity to tolerate high-intensity exercise.