An in vitro model of chronic wounding and its implication for age-related macular degeneration

Degeneration of the retinal pigment epithelium (RPE) plays a central role in age-related macular degeneration (AMD). Throughout life, RPE cells are challenged by a variety of cytotoxic stressors, some of which are cumulative with age and may ultimately contribute to drusen and lipofuscin accumulation. Stressors such as these continually damage RPE cells resulting in a state of chronic wounding. Current cell-based platforms that model a state of chronic RPE cell wounding are limited, and the RPE cellular response is not entirely understood. Here, we used the electric cell-substrate impedance sensing (ECIS) system to induce a state of acute or chronic wounding on differentiated human fetal RPE cells to analyze changes in the wound repair response. RPE cells surrounding the lesioned area employ both cell migration and proliferation to repair wounds but fail to reestablish their original cell morphology or density after repetitive wounding. Chronically wounded RPE cells develop phenotypic AMD characteristics such as loss of cuboidal morphology, enlarged size, and multinucleation. Transcriptomic analysis suggests a systemic misregulation of RPE cell functions in bystander cells, which are not directly adjacent to the wound. Genes associated with the major RPE cell functions (LRAT, MITF, RDH11) significantly downregulate after wounding, in addition to differential expression of genes associated with the cell cycle (CDK1, CDC6, CDC20), inflammation (IL-18, CCL2), and apoptosis (FAS). Interestingly, repetitive wounding resulted in prolonged misregulation of genes, including FAS, LRAT, and PEDF. The use of ECIS to induce wounding resulted in an over-representation of AMD-associated genes among those dysregulated genes, particularly genes associated with advanced AMD. This simple system provides a new model for further investigation of RPE cell wound response in AMD pathogenesis

Influence of beetroot juice supplementation on intermittent exercise performance.

This is the final version of the article. Available from Springer on open access via the DOI in this record.PURPOSE: This study tested the hypothesis that nitrate (NO3 (-)) supplementation would improve performance during high-intensity intermittent exercise featuring different work and recovery intervals. METHOD: Ten male team-sport players completed high-intensity intermittent cycling tests during separate 5-day supplementation periods with NO3 (-)-rich beetroot juice (BR; 8.2 mmol NO3 (-) day(-1)) and NO3 (-)-depleted beetroot juice (PL; 0.08 mmol NO3 (-) day(-1)). Subjects completed: twenty-four 6-s all-out sprints interspersed with 24 s of recovery (24 × 6-s); seven 30-s all-out sprints interspersed with 240 s of recovery (7 × 30-s); and six 60-s self-paced maximal efforts interspersed with 60 s of recovery (6 × 60-s); on days 3, 4, and 5 of supplementation, respectively. RESULT: Plasma [NO2 (-)] was 237 % greater in the BR trials. Mean power output was significantly greater with BR relative to PL in the 24 × 6-s protocol (568 ± 136 vs. 539 ± 136 W; P  0.05). The increase in blood [lactate] across the 24 × 6-s and 7 × 30-s protocols was greater with BR (P  0.05). CONCLUSION: BR might be ergogenic during repeated bouts of short-duration maximal-intensity exercise interspersed with short recovery periods, but not necessarily during longer duration intervals or when a longer recovery duration is applied. These findings suggest that BR might have implications for performance enhancement during some types of intermittent exercise

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

PublishedJournal ArticleThis is the author accepted manuscript. The final version is available from American Physiological Society via the DOI in this record.We tested the hypothesis that dietary nitrate (NO3-)-rich beetroot juice (BR) supplementation could partially offset deteriorations in O2 transport and utilization and exercise tolerance after blood donation. Twenty-two healthy volunteers performed moderate-intensity and ramp incremental cycle exercise tests prior to and following withdrawal of ~450 ml of whole blood. Before donation, all subjects consumed seven 70-ml shots of NO3--depleted BR [placebo (PL)] in the 48 h preceding the exercise tests. During the 48 h after blood donation, subjects consumed seven shots of BR (each containing 6.2 mmol of NO3-, n = 11) or PL (n. = 11) before repeating the exercise tests. Hemoglobin concentration and hematocrit were reduced by ~8-9% following blood donation (P < 0.05), with no difference between the BR and PL groups. Steady-state 02 uptake during moderate-intensity exercise was ~4% lower after than before donation in the BR group (P < 0.05) but was unchanged in the PL group. The ramp test peak power decreased from predonation (341 ± 70 and 331 ± 68 W in PL and BR, respectively) to postdonation (324 ± 69 and 322 ± 66 W in PL and BR, respectively) in both groups (P < 0.05). However, the decrement in performance was significantly less in the BR than PL group (2.7% vs. 5.0%, P < 0.05). NO3 supplementation reduced the 02 cost of moderate-inten-sity exercise and attenuated the decline in ramp incremental exercise performance following blood donation. These results have implications for improving functional capacity following blood loss.We thank James White Drinks (Ipswich, UK) for donating the juices used in the study. We also thank Matthew Black, James Kelly, and Daryl Wilkerson for assistance with data processing

Two weeks of watermelon juice supplementation improves nitric oxide bioavailability but not endurance exercise performance in humans

This is the author accepted manuscript. The final version is available from the publisher via the DOI in this record.This study tested the hypothesis that watermelon juice supplementation would improve nitric oxide bioavailability and exercise performance. Eight healthy recreationally-active adult males reported to the laboratory on two occasions for initial testing without dietary supplementation (control condition). Thereafter, participants were randomly assigned, in a cross-over experimental design, to receive 16 days of supplementation with 300 mL·day(-1) of a watermelon juice concentrate, which provided ∼3.4 g l-citrulline·day(-1) and an apple juice concentrate as a placebo. Participants reported to the laboratory on days 14 and 16 of supplementation to assess the effects of the interventions on blood pressure, plasma [l-citrulline], plasma [l-arginine], plasma [nitrite], muscle oxygenation and time-to-exhaustion during severe-intensity exercise. Compared to control and placebo, plasma [l-citrulline] (29 ± 4, 22 ± 6 and 101 ± 23 μM), [l-arginine] (74 ± 9, 67 ± 13 and 116 ± 9 μM) and [nitrite] (102 ± 29, 106 ± 21 and 201 ± 106 nM) were higher after watermelon juice supplementation (P < 0.01). However, systolic blood pressure was higher in the watermelon juice (130 ± 11) and placebo (131 ± 9) conditions compared to the control condition (124 ± 8 mmHg; P < 0.05). The skeletal muscle oxygenation index during moderate-intensity exercise was greater in the watermelon juice condition than the placebo and control conditions (P < 0.05), but time-to-exhaustion during the severe-intensity exercise test (control: 478 ± 80, placebo: 539 ± 108, watermelon juice: 550 ± 143 s) was not significantly different between conditions (P < 0.05). In conclusion, while watermelon juice supplementation increased baseline plasma [nitrite] and improved muscle oxygenation during moderate-intensity exercise, it increased resting blood pressure and did not improve time-to-exhaustion during severe-intensity exercise. These findings do not support the use of watermelon juice supplementation as a nutritional intervention to lower blood pressure or improve endurance exercise performance in healthy adults