Quantification of complex DNA damage by ionising radiation: An experimental and theoretical approach

This thesis was submitted for the degree of Doctor of Philosophy and awarded by Brunel University.Ionising radiation potentially produces a broad spectrum of damage in DNA including single and double strand breaks (ssb and dsb) and base damages. It has been hypothesised that sites of complex damage within cellular DNA have particular biological significance due to an associated decreased efficiency in repair. The aim of this study is to gain further understanding of the formation of complex DNA damage. Irradiations of plasmid DNA illustrate that an increase in ionising density of the radiation results in a decrease in ssb yields/Gy but an increase in dsb per ssb, indicative of an increase in the number of complex damage sites per simple isolated damage site. As the mechanism for damage formation shifts from purely indirect at low scavenging capacities to a significant proportion of direct at higher scavenging capacities the proportion of complex damage increases. Comparisons with the yields of ssb and dsb simulated by Monte-Carlo calculations for AIK USX and a-particles also indicate this correspondence. The ionisation density of low energy, secondary electrons produced by photons was assessed experimentally from the dependence of the yield of OH radicals escaping intra-track recombination on photon energy. As energy decreases the OH radical yield initially decreases reflecting an increased ionisation density. However, with further decrease in photon energy the yield of OH radicals increases in line with theoretical calculations. Base damage yields were determined for low and high ionising density radiation over a range of scavenging capacities. As scavenging capacity increases the base damage:ssb ratios increases implying a contribution from electrons to base damage. It is proposed that base damage contributes to DNA damage complexity. Complex damage analysis reveals that at cell mimetic scavenging capacities, 23% and 72% of ssb have an additional spatially close damage site following y-ray and a-particle irradiation respectively.Commission of the European Community (Contract No. F14-CT95-0011c

Gastric Emptying Rate of Young Cyclists by Magnetic Resonance Imaging: Three Case Studies

Gastric emptying rate during exercise has been documented in adult athletes, but not in younger athletes because of the utilization of invasive techniques. However, magnetic resonance imaging (MRI) is a noninvasive alternative. The aim of this study was to pilot the sequencing of MRI to a typical fluid intake strategy, in order to assess the viability of the procedure. Therefore, we compared the gastric emptying rate of water and a carbohydrate drink in three well-trained 17-year-old cyclists using MRI. Each participant cycled 4 bouts of 15 minutes each at moderate intensity followed by a ∼15-minute time trial. Water or carbohydrate drinks were distributed in amounts of 200–250 mL. After an initial baseline MRI scan, cyclists ingested the first drink, cycled 15 minutes, followed by an MRI scan to assess remaining gastric volume. This feeding-exercise-scanning protocol was repeated thereafter. Relative to the initial volume at the start of the time trial, 69% and 72% of water and carbohydrate emptied during the time trial. MRI appears to be a feasible technique to assess gastric emptying rates in youth cyclists

Enhanced task-related brain activation and resting perfusion in healthy older adults after chronic blueberry supplementation

© 2017, Canadian Science Publishing. All rights reserved. Blueberries are rich in flavonoids, which possess antioxidant and anti-inflammatory properties. High flavonoid intakes attenuate age-related cognitive decline, but data from human intervention studies are sparse. We investigated whether 12 weeks of blueberry concentrate supplementation improved brain perfusion, task-related activation, and cognitive function in healthy older adults. Participants were randomised to consume either 30 mL blueberry concentrate providing 387 mg anthocyanidins (5 female, 7 male; age 67.5 ± 3.0 y; body mass index, 25.9 ± 3.3 kg·m−2) or isoenergetic placebo (8 female, 6 male; age 69.0 ± 3.3 y; body mass index, 27.1 ± 4.0 kg·m−2). Pre-and postsupplementation, participants undertook a battery of cognitive function tests and a numerical Stroop test within a 1.5T magnetic resonance imaging scanner while functional magnetic resonance images were continuously acquired. Quantitative resting brain perfusion was determined using an arterial spin labelling technique, and blood biomarkers of inflammation and oxidative stress were measured. Significant increases in brain activity were observed in response to blueberry supplementation relative to the placebo group within Brodmann areas 4/6/10/ 21/40/44/45, precuneus, anterior cingulate, and insula/thalamus (p < 0.001) as well as significant improvements in grey matter perfusion in the parietal (5.0 ± 1.8 vs-2.9 ± 2.4%, p = 0.013) and occipital (8.0 ± 2.6 vs-0.7 ± 3.2%, p = 0.031) lobes. There was also evidence suggesting improvement in working memory (2-back test) after blueberry versus placebo supplementation (p = 0.05). Supplementation with an anthocyanin-rich blueberry concentrate improved brain perfusion and activation in brain areas associated with cognitive function in healthy older adults

Dietary nitrate supplementation attenuates the reduction in exercise tolerance following blood donation

Dietary nitrate supplementation attenuates the reduction in exercise tolerance following blood donatio

Contralateral fatigue during severe-intensity single-leg exercise: influence of acute acetaminophen ingestion

Exhaustive single-leg exercise has been suggested to reduce time to task failure (Tlim) during subsequent exercise in the contralateral leg by exacerbating central fatigue development. We investigated the influence of acetaminophen (ACT), an analgesic which may blunt central fatigue development, on Tlim during single-leg exercise completed both with, and without, prior fatiguing exercise of the contralateral leg. Fourteen recreationally-active men performed single-leg, severe-intensity knee extensor exercise to Tlim on the left (Leg1) and right (Leg2) legs without prior contralateral fatigue, and on Leg2 immediately following Leg1 (Leg2- CONTRA). The tests were completed following ingestion of 1 g ACT or maltodextrin (placebo) capsules. Intramuscular phosphorous-containing metabolites and substrates, and muscle activation, were assessed using 31 P-MRS and electromyography, respectively. Tlim was not different between the Leg1ACT and Leg1PL conditions (402 ± 101 vs. 390 ± 106 s; P=0.11). There was also no difference in Tlim between Leg2ACT-CONTRA and Leg2PL-CONTRA (324 ± 85 vs. 311 ± 92 s; P=0.10), but Tlim was shorter in these tests compared to Leg2CON (385 ± 104 s; both P<0.05). There were no differences in intramuscular phosphorous-containing metabolites and substrates, or muscle activation, between the Leg1ACT and Leg1PL or the Leg2ACT-CONTRA and Leg2PL-CONTRA conditions (all P>0.05). These findings suggest that levels of metabolic perturbation and muscle activation are not different at task failure during single leg severe-intensity knee extensor exercise completed with or without prior fatiguing exercise of the contralateral leg. Despite the existence of contralateral fatigue, ACT ingestion did not alter neuromuscular responses or exercise performance

The effects of β-alanine supplementation on muscle pH and the power-duration relationship during high-intensity exercise

Purpose: To investigate the influence of β-alanine (BA) supplementation on muscle carnosine content, muscle pH and the power-duration relationship (i.e., critical power and W′). Methods: In a double-blind, randomized, placebo-controlled study, 20 recreationally-active males (22 ± 3 y, VO˙2peak 3.73 ± 0.44 L·min−1) ingested either BA (6.4 g/d for 28 d) or placebo (PL) (6.4 g/d) for 28 d. Subjects completed an incremental test and two 3-min all-out tests separated by 1-min on a cycle ergometer pre- and post-supplementation. Muscle pH was assessed using 31P-magnetic resonance spectroscopy (MRS) during incremental (INC KEE) and intermittent knee-extension exercise (INT KEE). Muscle carnosine content was determined using 1H-MRS. Results: There were no differences in the change in muscle carnosine content from pre- to post-intervention (PL: 1 ± 16% vs. BA: −4 ± 25%) or in muscle pH during INC KEE or INT KEE (P > 0.05) between PL and BA, but blood pH (PL: −0.06 ± 0.10 vs. BA: 0.09 ± 0.13) during the incremental test was elevated post-supplementation in the BA group only (P < 0.05). The changes from pre- to post-supplementation in critical power (PL: −8 ± 18 W vs. BA: −6 ± 17 W) and W′ (PL: 1.8 ± 3.3 kJ vs. BA: 1.5 ± 1.7 kJ) were not different between groups. No relationships were detected between muscle carnosine content and indices of exercise performance. Conclusions: BA supplementation had no significant effect on muscle carnosine content and no influence on intramuscular pH during incremental or high-intensity intermittent knee-extension exercise. The small increase in blood pH following BA supplementation was not sufficient to significantly alter the power-duration relationship or exercise performance

Network analysis of nitrate-sensitive oral microbiome reveals interactions with cognitive function and cardiovascular health across dietary interventions

Many oral bacteria reduce inorganic nitrate, a natural part of a vegetable-rich diet, into nitrite that acts as a precursor to nitric oxide, a regulator of vascular tone and neurotransmission. Aging is hallmarked by reduced nitric oxide production with associated detriments to cardiovascular and cognitive function. This study applied a systems-level bacterial co-occurrence network analysis across 10-day dietary nitrate and placebo interventions to test the stability of relationships between physiological and cognitive traits and clusters of co-occurring oral bacteria in older people. Relative abundances of Proteobacteria increased, while Bacteroidetes, Firmicutes and Fusobacteria decreased after nitrate supplementation. Two distinct microbiome modules of co-occurring bacteria, that were sensitive to nitrate supplementation, showed stable relationships with cardiovascular (Rothia-Streptococcus) and cognitive (Neisseria-Haemophilus) indices of health across both dietary conditions. A microbiome module (Prevotella-Veillonella) that has been associated with pro-inflammatory metabolism was diminished after nitrate supplementation, including a decrease in relative abundance of pathogenic Clostridium difficile. These nitrate-sensitive oral microbiome modules are proposed as potential pre- and probiotic targets to ameliorate age-induced impairments in cardiovascular and cognitive health.publishedVersio

Montmorency cherry supplementation attenuates vascular dysfunction induced by prolonged forearm occlusion in overweight, middle-aged men

Flavonoid supplementation improves brachial artery flow-mediated dilation (FMD), but it is not known whether flavonoids protect against vascular dysfunction induced by ischemia-reperfusion (IR) injury and associated respiratory burst. In a randomized, double-blind, placebo-controlled, crossover study, we investigated whether 4 wk supplementation with freeze-dried Montmorency cherry (MC) attenuated suppression of FMD after IR induced by prolonged forearm occlusion. Twelve physically inactive overweight, middle-aged men (52.8 ± 5.8 yr, BMI: 28.1 ± 5.3 kg/m2) consumed MC (235 mg/day anthocyanins) or placebo capsules for 4 wk, with supplementation blocks separated by 4 wk washout. Before and after each supplementation block, FMD responses and plasma nitrate and nitrite ([ N O − 2 ]) concentrations were measured at baseline and 15, 30, and 45 min after prolonged (20 min) forearm occlusion. FMD response was significantly depressed by the prolonged occlusion ( P < 0.001). After a 45-min reperfusion, FMD was restored to baseline levels after MC (ΔFMD presupplementation: -30.5 ± 8.4%, postsupplementation: -0.6 ± 9.5%) but not placebo supplementation (ΔFMD presupplementation: -11.6 ± 10.6, postsupplementation: -25.4 ± 4.0%; condition × supplement interaction: P = 0.038). Plasma [ N O − 2 ] decreased after prolonged occlusion but recovered faster after MC compared with placebo (Δ45 min to baseline; MC: presupplementation: -15.3 ± 9.6, postsupplementation: -6.2 ± 8.1; Placebo: presupplementation: -16.3 ± 5.9, postsupplementation: -27.7 ± 11.1 nmol/l; condition × supplement × time interaction: P = 0.033). Plasma peroxiredoxin concentration ([Prx2]) was significantly higher after MC (presupplementation: 22.8 ± 1.4, postsupplementation: 28.0 ± 2.4 ng/ml, P = 0.029) but not after placebo supplementation (presupplementation: 22.1 ± 2.2, postsupplementation: 23.7 ± 1.5 ng/ml). In conclusion, 4 wk MC supplementation enhanced recovery of endothelium-dependent vasodilatation after IR, in parallel with faster recovery of plasma [ N O − 2 ], suggesting NO dependency. These protective effects seem to be related to increased plasma [Prx2], presumably conferring protection against the respiratory burst during reperfusion. NEW & NOTEWORTHY This is the first study to demonstrate that 4 wk of Montmorency cherry powder supplementation exerted protective effects on endothelium-dependent vasodilation after transient ischemia-reperfusion injury in overweight, physically inactive, nonmedicated, hypertensive middle-aged men. These effects seem to be due to increased nitric oxide availability, as evidenced by higher plasma nitrite concentration and peak arterial diameter during the flow-mediated dilation measurement. This may be a consequence of increased concentration of peroxiredoxin and other antioxidant systems and, hence, reduced reactive oxygen species exposure.This article is freely available via Open Access. Click on the Publisher URL to access it via the publisher's site.This study was partially funded by a grant from the Cherry Research Committee, Aboo-Bakkar was supported by a Ph.D. studentship from the Universiti Kuala Lumpur, and Fulford’s salary was provided by National Institute for Health Research.accepted version (12 month embargo

Dietary nitrate supplementation improves sprint and high-intensity intermittent running performance

Dietary nitrate supplementation improves sprint and high-intensity intermittent running performanc

Discrete physiological effects of beetroot juice and potassium nitrate supplementation following 4 weeks sprint interval training

The physiological and exercise performance adaptations to sprint interval training (SIT) may be modified by dietary nitrate (NO3-) supplementation. However, it is possible that different types of NO3- supplementation evoke divergent physiological and performance adaptations to SIT. The purpose of this study was to compare the effects of 4 weeks SIT with and without concurrent dietary NO3- supplementation administered as either NO3--rich beetroot juice (BR) or potassium NO3- (KNO3). Thirty recreationally-active subjects completed a battery of exercise tests before and after a 4 week intervention in which they were allocated to one of three groups: 1) SIT undertaken without dietary NO3- supplementation (SIT); 2) SIT accompanied by concurrent BR supplementation (SIT+BR); or 3) SIT accompanied by concurrent KNO3 supplementation (SIT+KNO3). During severe-intensity exercise, VO2peak and time to task failure were improved to a greater extent with SIT+BR than SIT and SIT+KNO3 (P0.05). These findings indicate that 4 weeks SIT with concurrent BR supplementation results in greater exercise capacity adaptations compared to SIT alone and SIT with concurrent KNO3 supplementation. This may be the result of greater NO-mediated signalling in SIT+BR compared to SIT+KNO3