Contribution Of Glycolysis Research Articles

An alternate to accumulated oxygen deficit (AOD) for measuring anaerobic contribution: 'AOD_alt' is valid in normoxia and hypoxia.

The gold standard measure of anaerobic contribution is accumulated oxygen deficit (AOD). The purpose of this study was to investigate the validity of an alternate measure, AOD_alt. AOD_alt is the sum of the phosphocreatine and glycolytic contributions, which are estimated from post-exercise oxygen uptake and blood lactate concentration, respectively. In Study One, six women and three men performed 6-min bouts of heavy intensity cycle ergometer exercise, once in normoxia (FIO2 ~ 21%) and twice under hypoxic conditions (FIO2 ~ 15% and ~ 12%). In Study Two, four women and two men performed severe intensity tests to exhaustion, once in normoxia (~ 10min) and twice in hypoxia (FIO2 ~ 15% and ~ 10%). Physiological responses were measured during exercise and 7min of recovery. In 6min of heavy exercise, Study One, the alternate and criterion measures of anaerobic contribution (AOD_alt and AOD) were correlated, in normoxia and in hypoxia. In exhaustive severe exercise, Study Two, AOD_alt and AOD were correlated (r = 0.77) and similar, in normoxia and at FIO2 ~ 15%. However, AOD_alt and AOD values were neither correlated (r = 0.27) nor similar (57 ± 5mL·kg-1 vs 51 ± 7mL·kg-1) at FIO2 ~ 10%. These results confirm the validity of AOD_alt as a measure of anaerobic capacity in severe intensity exercise, demonstrate its validity in heavy exercise, and assert its validity in conditions of hypoxia (FIO2 ~ 12%).

A 6-Week Ketogenic Diet Enhances the Phosphocreatine Energy System Contribution During Intermittent Sprints

Abstract Purpose Team sports often involve intermittent sprints. During these activities the Phosphocreatine-ATP buffer (ATP-PCr) signifies the major anaerobic energy substrate. While the effects of ketogenic diets (KD) on carbohydrate and fat metabolism during endurance exercise are widely reported, we explored keto-adaptation in ATP-PCr metabolism during intermittent sprint exercise. Methods Following a within-subject repeated measures design, 15 recreationally active participants (7 men, 8 women, aged 25.1 ± 6.4 years) performed cycle ergometer intermittent sprints (6 × 10 s sprints, 2 min recovery) with VO2 and blood lactate measurements for energy system calculations. These laboratory tests were performed in alternate weeks; First, twice at baseline on their habitual diet (HD) (35% CHO, 45% fat, 20% protein) and thereafter over a 6-week KD (7% CHO, 66% fat, 28% protein). Results Repeated measures ANOVA’s and Bonferroni tests revealed ATP-PCr derived energy increased significantly from HD to KD week 6 (+ 22.0 ± 43.15 J; P = 0.019; ES = 0.47). From HD to KD week 2, anaerobic glycolytic contribution lowered (− 14.4 ± 28.16 J; P = 0.031; ES = − 0.10) and peak blood [lactate] reduced significantly (− 2.92 ± 0.851 mmol; P = 0.004; ES = − 0.73). There was no statistically significant within-subject change in mean sprint power (P = 0.356). Conclusion The 6-week KD did not compromise intermittent sprint performance. The findings suggest that the ATP-PCr energy pathway may be a novel site of metabolic keto-adaptation. This, combined with the lowered blood [lactate] we observed, presents desirable metabolic adaptations for intermittent sprint sport athletes.

Calculation of a conversion factor for estimating the glycolytic contribution in exercise from post-exercise blood lactate concentration.

Purpose: Often, the glycolytic contribution in a bout of heavy or severe intensity exercise is estimated by multiplying the increase in blood lactate concentration above resting levels that is engendered by the exercise (in mM) by 3.3 (or 3) mL·kg-1 per mM. Our purpose was to verify the value of this conversion factor, using methods that were completely different from those of the original studies. Methods: Six women (mean ± SD), age, 23 ± 1year; VO2max, 46 ± 4mL·kg-1·min-1) and three men (23 ± 0years; 54 ± 8mL·kg-1·min-1) completed 6min of heavy intensity exercise in conditions of normoxia and hypoxia (FIO2, ∼12%). VO2 was measured throughout the exercise and 7min of recovery. The increase in glycolytic contribution was estimated as the reduction in aerobic contribution in hypoxia, after correction for the effects of hypoxia on the oxygen demand and on the contribution from phosphocreatine. The peak post-exercise blood lactate concentration was measured in fingerstick blood samples. Results: The ratio between the increase in estimated glycolytic contribution (in mL·kg-1) in hypoxia and the increase in peak blood lactate concentration (in mM) yielded an oxygen equivalent of 3.4 ± 0.4mL·kg-1 per mM (range, 2.6mL·kg-1 per mM to 4.0mL·kg-1 per mM) for cycle ergometer exercise. Conclusion: These results generally support the use of a common conversion factor to calculate the glycolytic contribution from post-exercise blood lactate concentrations. However, there is some inter-individual variability in the conversion factor.

Energy System Contributions during Olympic Combat Sports: A Narrative Review.

This narrative review focuses on the studies that estimate the energy systems' contributions during match simulations of striking (boxing, karate, and taekwondo), grappling (judo), and weapon-based (fencing) Olympic combat sports. The purpose is to provide insights into the metabolism of these athletes. In striking Olympic combat sports, the oxidative contribution varied from 62% (in karate and taekwondo) to 86% (in boxing), the ATP-PCr system contribution varied from 10% (in boxing) to 31% (in taekwondo), and the glycolytic contribution was between 3% (in the third round of taekwondo) and 21% (in karate). In grappling combat sports, only judo was studied, and for a 4 min match, the oxidative contribution was 79%, followed by 14% ATP-PCr system contribution and 7% contribution from the glycolytic system. In fencing, the only weapon-based Olympic combat sport, the oxidative contribution varied from 81% (in the first bout) to 90% (in the second bout), followed by 9% (bout 2) to 12% (bout 1) contribution from the ATP-PCr system, and 0.6% to 7% contribution from the glycolytic system during 3 × 3 min bouts of épée match simulation. Hence, Olympic combat sports are primarily powered by the oxidative system, but the key scoring actions are likely fueled by anaerobic pathways.

The Oxidative-Glycolytic Balance Influenced by Sprint Duration Is Key during Repeated Sprint in Hypoxia.

This study investigates the effects of normobaric hypoxia on repeated sprint exercise (RSE) with different balance between oxidative (phosphocreatine and oxidative pathway) and glycolytic contributions. Therefore, performance and psychophysiological responses were compared during RSE to exhaustion with the same exercise-to-rest ratio (1:2) but different sprint durations (5, 10, or 20 s) either in normoxic (RSN) or hypoxic (RSH; F io2 = 0.13) conditions. On separate visits, 10 active participants completed in random order three cycling RSN (5:10; 10:20 and 20:40) and three similar RSH sessions to exhaustion. Vastus lateralis muscle oxygenation was recorded by near-infrared spectroscopy. Blood lactate concentration, limb and breathing discomfort, and ratings of perceived exertion were measured. Total sprint number was smaller in hypoxia than in normoxia for 5:10 (20.8 ± 8.6 vs 14.7 ± 3.4; P = 0.014) and 10:20 (13.7 ± 6.3 vs 8.8 ± 2.5; P = 0.018) but not 20:40 (5.6 ± 1.9 vs 5.6 ± 2.5). The fatigue index was larger in hypoxia only for 5:10 (-43.5%, P < 0.001). Irrespective of condition, blood lactate concentration increased with the sprint duration with higher values for 20:40 than 5:10 (13.1 ± 2.7 vs 11.5 ± 2.2 mmoL·L -1 ; P = 0.027). Limb and breathing discomfort and ratings of perceived exertion did not differ in all RSE. Muscle oxygenation was mainly impacted by sprint duration (i.e., main effect of sprint duration on [HHb] min, [tHb] max, Δ[HHb], and Δ[tHb]) but not by hypoxia. The normoxia-to-hypoxia percentage decrease for total sprint number for 5:10 was correlated with the highest power output over 5 s ( R2 = 0.55; P = 0.013) and 10 s ( R2 = 0.53; P = 0.016). Hypoxia impairs repeated sprint ability when the oxidative but not the glycolytic contribution is substantial. The oxidative-glycolytic balance, influenced partly by sprint duration, is key during repeated sprint in hypoxia.

The underlying mechanisms of the effect of superchilling on the tenderness of beef Longissimus lumborum

This study investigated the effect of superchilling (−30 °C until the core temperature achieved −3 °C, then stored at −1 °C until 24 h, SC) on the tenderness of hot boned beef M. longissimus lumborum (LL), with very fast chilling (−30 °C until the core temperature achieved 0 °C, then stored at −1 °C until 24 h, VFC) and conventional chilling (0– 4 °C for 24 h, CC) as the controls. The lowest initial shear force values were obtained in SC samples compared to those from the VFC and CC treatments (P < 0.05). Clear freezing damage of muscle fibers and more myofibril fragmentation were found in SC samples compared with the other samples early post-mortem. Moreover, SC samples showed the highest level of inosine 5-monophosphate at 3 h post-mortem (P < 0.05). A reduced glycolysis rate (as evidenced by lactate content) was also found in SC treated samples suggesting little contribution of glycolysis on the tenderization of SC.

Physiological Profiling and Energy System Contributions During Simulated Epée Matches in Elite Fencers.

The aim of this study was to investigate physiological responses and energetic contributions during simulated epée matches in elite fencers. Ten elite male fencers participated in simulated epée (direct elimination) matches. Simulated epée matches included 3 bouts of 3minutes each with 1-minute rests between bouts. During these sessions, physiological variables such as mean and peak heart rate, peak and mean oxygen uptake (VO2peak and VO2mean), metabolic equivalents of task in VO2peak and VO2mean, and blood lactate concentrations (peak lactate concentration and delta blood lactate concentration) were measured. Furthermore, energetic contributions (oxidative [WOxi], glycolytic, and phosphagen) and time-motion variables were estimated. Values of peak heart rate, mean heart rate, and WOxi (in percentages) were significantly higher in the second and third bouts compared with the first. VO2peak and metabolic equivalents of task in VO2peak were significantly higher in the first bout compared with the third bout. Values of delta blood lactate concentration and glycolytic contribution (in kilojoules and percentages) were significantly lower in the second and third bouts compared with the first. VO2mean and metabolic equivalents of task in VO2mean were significantly higher in the second bout compared with the third bout. Furthermore, WOxi (in kilojoules and percentage) was significantly higher in all bouts compared with phosphagen and glycolytic contributions. Low positive and negative correlations were seen between WOxi, VO2mean, sum of attacks and defense times, and the sum of time without attacks and defenses. Direct-elimination epée matches consist of high-intensity intermittent exercise, and the oxidative contribution is 80% to 90%. Improving aerobic conditioning may support high-intensity intermittent actions during entire epée matches in elite fencers.

Does Inosine Monophosphate Formation Facilitate the Bioenergetic Response to Incremental Contractions in Human Skeletal Muscle In Vivo?

Traditionally, it has been accepted that working muscle will endeavor to maintain [ATP] near its resting level in order to maintain a favorable free energy of ATP hydrolysis (∆GATP). Recent reports of significant declines in [ATP] during maximal muscular contractions in vivo spark new interest in this concept. ATP is hydrolyzed to adenosine diphosphate (ADP) in the creatine kinase reaction to provide energy directly to working ATPases. As needed, additional energy can be provided by the breakdown of ADP to adenosine monophosphate (AMP), which is rapidly cleared by the formation of inosine monophosphate (IMP), with accompanying loss of total adenine nucleotides. IMP formation has been thought to occur only during very intense muscular work, wherein ATP production does not keep pace with ATP consumption. Here, we propose a role for IMP in managing both ∆GATP and flux through a key energetic pathway during contractions. To test our hypothesis that IMP formation mitigates the accumulation of ADP, 8 males (28.4 + 3.5 yr, mean + SD) performed 8 min of incremental, isotonic knee extension contractions (1 every 2 s, starting at 6% peak torque and incrementing by 3% every 2 min) on a magnetic resonance (MR)‐compatible ergometer in a 3‐Tesla MR system. 31Phosphorus MR spectra were obtained at rest (60‐s average) and throughout the contraction protocol (10‐s temporal resolution). Peaks corresponding to ATP, phosphocreatine (PCr), inorganic phosphate (Pi), and phosphomonoesters (PME) were fit in jMRUI, and concentrations for these metabolites were determined assuming resting [ATP] = 8.2 mM. [ADP], [AMP], [IMP], and the rate of ATP (mM∙s‐1) production by non‐oxidative glycolysis (i.e., wherein the fate of pyruvate is lactate; ATPgly) were also calculated. As expected, [PCr] decreased and [Pi] increased approximately linearly throughout the incremental contraction protocol. Surprisingly, [ATP] decreased and [IMP] increased (to 7.3 + 0.4 mM and 0.8 + 0.56 mM, respectively) steadily from the onset of contractions until the final stage, at which point no further changes were observed. In contrast, both [ADP] and [AMP] increased linearly from the start of contractions until the final stage, when they accumulated more rapidly; ultimately to 109.9 + 52.1 uM and 1.0 + 0.91 uM, respectively. The glycolytic contribution to ATP production was consistent for the first 3 stages, after which ATPgly increased rapidly to 0.102 + 0.14 mM·s‐1 at 480 s. Collectively, the timing and magnitude of these metabolic changes in working muscle suggest a sequence of responses designed to maintain cytosolic [ADP] sufficiently low as to mitigate changes in ∆GATP, but at a level sufficient to stimulate ATPgly as contraction intensity increases. A novel observation in this study was the decrease in [ATP] (and thus increase in [IMP]) from the onset of the contraction protocol, which then leveled off at the highest contraction intensity. The utility and consequences of these responses remain to be determined, but this study supports the concept that IMP formation is important in maintaining the cellular energy state as well as in directing the bioenergetic response to a series of incremental muscular contractions in vivo.

Effects of Oral Creatine Supplementation on Power Output during Repeated Treadmill Sprinting.

The aim of the present study was to examine the effects of creatine (Cr) supplementation on power output during repeated sprints on a non-motorized treadmill. Sixteen recreationally active males volunteered for this study (age 25.5 ± 4.8 y, height 179 ± 5 cm, body mass 74.8 ± 6.8 kg). All participants received placebo supplementation (75 mg of glucose·kg−1·day−1) for 5 days and then performed a baseline repeated sprints test (6 × 10 s sprints on a non-motorised treadmill). Thereafter, they were randomly assigned into a Cr (75 mg of Cr monohydrate·kg−1·day−1) or placebo supplementation, as above, and the repeated sprints test was repeated. After Cr supplementation, body mass was increased by 0.99 ± 0.83 kg (p = 0.007), peak power output and peak running speed remained unchanged throughout the test in both groups, while the mean power output and mean running speed during the last 5 s of the sprints increased by 4.5% (p = 0.005) and 4.2% to 7.0%, respectively, during the last three sprints (p = 0.005 to 0.001). The reduction in speed within each sprint was also blunted by 16.2% (p = 0.003) following Cr supplementation. Plasma ammonia decreased by 20.1% (p = 0.037) after Cr supplementation, despite the increase in performance. VO2 and blood lactate during the repeated sprints test remained unchanged after supplementation, suggesting no alteration of aerobic or glycolytic contribution to adenosine triphosphate production. In conclusion, Cr supplementation improved the mean power and speed in the second half of a repeated sprint running protocol, despite the increased body mass. This improvement was due to the higher power output and running speed in the last 5 s of each 10 s sprint.

High-intensity resistance exercise is not as effective as traditional high-intensity interval exercise for increasing the cardiorespiratory response and energy expenditure in recreationally active subjects

PurposeTraditional high-intensity interval exercise (HIIE) highly stimulates the cardiorespiratory system and increases energy expenditure (EE) during exercise. High-intensity resistance exercise (HIRE) has become more popular in recreationally active subjects. The physiological responses to HIRE performed with light or moderate load is currently largely unknown. Here, we examined the effect of the type of interval exercise [HIRE at 40% (HIRE40) and 60% (HIRE60) 1-RM vs. traditional HIIE] on the cardiorespiratory response and EE during and after exercise.MethodsFifteen recreationally active adults randomly completed traditional HIIE on an ergocyle, HIRE40 and HIRE60. The sessions consisted of two sets of ten 30-s intervals (power at 100% VO2max during HIIE; maximal number of repetitions for 10 different free-weight exercises during HIRE40 and HIRE60) separated by 30-s active recovery periods. Gas exchange, heart rate (HR) and EE were assessed during and after exercise.ResultsVO2mean, VO2peak, HRmean, the time spent above 90% VO2max and HRmax, and aerobic EE were lower in both HIRE sessions compared with HIIE (P < 0.05). Anaerobic glycolytic contribution to total exercise EE was higher in HIRE40 and HIRE60 compared with HIIE (P < 0.001). EE from excess post-exercise oxygen consumption (EPOC) was similar after the three sessions. Overall, similar cardiorespiratory responses and EE were found in HIRE40 and HIRE60.ConclusionsHIRE is not as effective as HIIE for increasing the cardiorespiratory response and EE during exercise, while EPOC remains similar in HIRE and HIIE. These parameters are not substantially different between HIRE40 and HIRE60.

Energetics contribution during no-gi Brazilian jiu jitsu sparring and its association with regional body composition.

We used measurements of metabolic perturbation obtained after sparring to estimate energetics contribution during no-gi Brazilian jiu-jitsu. Ten advanced grapplers performed two six-minute sparring bouts separated by 24 hours. Kinetics of recovery rate of oxygen uptake was modelled and post-combat-sparring blood-lactate concentration measured to estimate oxygen equivalents for phospholytic and glycolytic components of anaerobic energetics, respectively. Linear regression was used to estimate end-combat-sparring rate of oxygen uptake. Regional and whole-body composition were assessed using dual X-ray absorptiometry with associations between these measurements and energy turnover explored using Pearson’s correlation coefficient (significance, P < 0.05). Estimated oxygen equivalents for phospholytic and glycolytic contributions to anaerobic metabolism were 16.9 ± 8.4 (~28%) and 44.6 ± 13.5 (~72%) mL∙kg-1, respectively. Estimated end-exercise rate of oxygen uptake was 44.2 ± 7.0 mL∙kg-1∙min-1. Trunk lean mass was positively correlated with both total anaerobic and glycolytic-specific energetics (total, R = 0.645, p = 0.044; glycolytic, R = 0.692, p = 0.027) and negatively correlated with end-exercise rate of oxygen uptake (R = -0.650, p = 0.042). There were no correlations for any measurement of body composition and phospholytic-specific energetics. Six minutes of no-gi Brazilian jiu-jitsu sparring involves high relative contribution from the glycolytic component to total anaerobic energy provision and the link between this energetics profile and trunk lean mass is consistent with the predominance of ground-based combat that is unique for this combat sport. Training programs for Brazilian jiu-jitsu practitioners should be designed with consideration given to these specific energetics characteristics.

Abstract 2332: Modifying bioavailability of ATP and measuring metabolic response in HeLa cells

Abstract Introduction: Since studies by Otto Warburg demonstrated that cancer cells had markedly reduced oxidative phosphorylation (OXPHOS) under conditions of high oxygen tension (Warburg Effect/aerobic glycolysis), metabolic reprogramming has become a fundamental hallmark of cancer. While numerous discoveries have revealed highly specific metabolic differences between cancer cells, utilization of aerobic glycolysis as the primary means of ATP generation remains one of the most common metabolic alterations observed across a range of cancers. There is growing interest in understanding the multifaceted role ATP plays in cancer development, progression and chemotherapeutic resistance. Using the man-made protein DX, which chelates ATP with high specificity and affinity, we have begun to untangle these roles by significantly reducing cytosolic ATP and investigating cellular response. Hypothesis: Delivery of DX into HeLa cancer cells will reduce intracellular bioavailable ATP and induce an adaptive metabolic response. Methods: A cationic lipid mixture was complexed with active, purified DX protein to generate a DX/lipid complex capable of delivering the protein to the cytoplasm of HeLa cells. To confirm the successful delivery of DX, immunofluorescent microscopy was performed (DYKDDDDK tag monoclonal antibody, Alexa Fluor 647) at various time points following DX transfection. The impact of DX on cell viability was determined using a tetrazolium-based colorimetric cell viability assay and a caspase 3/7 assay. To correlate phenotypic/viability change with DX activity, bioavailable ATP levels were measured at specific time points following DX delivery. Additionally, the relative contribution of glycolysis and OXPHOS to the total ATP production rate was measured using the label-free XF Real Time ATP Rate Assay (XFe96 Seahorse, Agilent Technologies) over time post DX transfection. Results: In a time- and dose- dependent manner, DX negatively impacted cell growth and induced cell death via apoptosis, at a time concomitant with a decrease in bioavailable ATP. In response to DX over time, the total ATP production rates in HeLa cells significantly decreased. Importantly, this reduction in the rate of ATP production was associated with a metabolic program that was primarily glycolytic (70%) with a smaller contribution from OXPHOS (30%) 24 h post DX transfection. Conclusion: Advances in protein engineering have made it possible to create artificial proteins with specific functions. In addition to direct clinical applications, synthetic proteins can be developed into powerful experimental tools to resolve biological enigmas. Direct delivery of DX potentiated energy flux in HeLa cells in an unbiased manner, independent of metabolic pathway interference. This work establishes DX as a useful tool for examination of cancer cell metabolism in response to ATP stress. Citation Format: Taha Muhammad, Ashley Brown, Selina Martinez, John C. Chaput, Jeffrey Norris, Shaleen Korch. Modifying bioavailability of ATP and measuring metabolic response in HeLa cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2332.

Effects of sprint distance and repetition number on energy system contributions in soccer players

Background/objectiveThis study aims to compare the effect of sprint distance and repetition number on performance, physiological responses, and energy systems contributions. MethodsEighteen male university league soccer players (age: 19.9 ± 1.6 years, height: 177.9 ± 4.7 cm, body mass: 72.4 ± 6.3 kg, percentage body fat: 8.9 ± 1.8, training experience: 7.4 ± 1.6 years) completed two different repeated sprint protocols: 20 × 20 m (20 × 20) and 10 × 40 m (10 × 40) with 15s and 30s rest intervals, respectively. Oxygen uptake (VO2) were measured during the rest, exercise, and recovery phases. Rest and peak blood lactate concentrations were determined. Using VO2 and lactate values, the energy system contributions were calculated using a mono-exponential model and mathematical calculations. Energy systems contributions and total energy expenditure (TEE) were calculated both for the entire protocol (overall) and for the sprints only. ResultsRatings of perceived exertion (RPE), peak and mean heart rate (HR) responses were significantly higher in the 20 × 20 whereas lactate response was higher in the 10 × 40. TEE was similar between the 10 × 40 (586.3 ± 60.8 kJ) and 20 × 20 (595.6 ± 57.5 kJ). For overall estimations, the 10 × 40 and 20 × 20 presented similar results of oxidative (47.5 ± 5.4 vs 45.7 ± 5.1 kJ min−1) and phosphagen (44.7 ± 5.4 vs (42.9 ± 4.8 kJ min−1) systems contributions whereas glycolytic contribution was higher in the 10 × 40 (15.5 ± 2.2 vs 12.8 ± 2.3 kJ min−1). For sprints only estimation, the phosphagen (257.6 ± 31.5 vs 225.2 ± 28.2 kJ min−1), glycolytic (89.4 ± 13.4 vs 67.3 ± 12.5 kJ min−1), and oxidative (76.9 ± 6.9 vs 72.0 ± 7.9 2 kJ min−1) systems contributions were higher in the 10 × 40. ConclusionsAlthough HR and RPE responses were higher in the 20 × 20, phosphagen (during sprints) and glycolytic (during both sprints and overall protocol) were higher in the 10 × 40 protocol. Therefore, the 10 × 40 protocol seems more reasonable for developing or evaluating the anaerobic systems.

Energy pathway contributions during 60-second upper-body Wingate test in Greco-Roman wrestlers: intermittent versus single forms

ABSTRACT This study aimed to investigate the energy pathway contributions and physiological and performance responses between a 10 × 6-second intermittent sprint test (IST) and a 60-second single maximal test (SMT). Seventeen highly trained male Greco-Roman wrestlers participated in this study. Participants completed the 60-second upper-body Wingate tests, both intermittent and single forms. The contributions of the oxidative, glycolytic, and ATP-PCr pathways were estimated using mathematical methods based on lactate values and oxygen consumption kinetics of rest, exercise, and recovery phases. The main findings indicated that total energy expenditure (TEE) and the contribution of oxidative, glycolytic, and ATP-PCr pathways were 514 kJ, 45%, 11%, and 44% for IST (overall: sprints + rest intervals); 333 kJ, 14%, 17%, and 69% for IST (sprints only); and 159 kJ, 31%, 38%, and 31% for SMT, respectively. TEE and ATP-PCR pathway contributions were higher in the IST (both overall and sprint only), whereas glycolytic pathway contribution and delta lactate were higher in the SMT. Absolute oxidative contribution was similar, but relative oxidative contribution was higher in the SMT. Additionally, mean power was higher in the IST than SMT, whereas peak power, peak and mean heart rate, and ratings of perceived exertion were similar.

Brain Endothelial Cells: Metabolic Flux and Energy Metabolism.

The neurovascular unit (NVU) consists of multiple cell types including brain endothelial cells, pericytes, astrocytes, and neurons that function collectively to maintain homeostasis within the CNS microenvironment. As the principal barrier-forming component of the NVU, the endothelial cells perform an array of complex functions that require substantial energy resources. The principal metabolic pathways for producing ATP are glycolysis and mitochondrial oxidative phosphorylation. While previous studies have demonstrated that glycolysis is a primary pathway for most endothelial cells, details about the energy producing pathways of brain endothelial cells are not fully characterized. The contributions of glycolysis and mitochondrial respiration to energy metabolism are quantifiable using metabolic flux analysis that measures cellular oxygen consumption and acidification (proton production) in a closed microtiter plate format. ATP production rates are then calculated. The bioenergetics of the human brain microvascular endothelial cell line, hCMEC/D3, indicate that these cells exhibit relatively elevated rates of glycolytic flux and glycolytic ATP production, thus confirming their glycolytic nature even in the presence of abundant oxygen. Furthermore, energy producing pathways involving mitochondrial respiration are relatively low, although contributing significantly to total ATP production. Interestingly, the bioenergetics of the hCMEC/D3 cells are relatively similar to those of human primary brain microvascular endothelial cells (hBVECs). These findings allow a quantitative understanding of the bioenergetics of brain endothelial cells in a cultured and proliferative state and also provide a platform for comparative studies of disease states and conditions involving exposures to drugs or metabolic disruptors.

Effect of Red &amp;lt;i&amp;gt;Panax ginseng&amp;lt;/i&amp;gt; on Mitochondrial Dynamics and Bioenergetics in HaCaT Cells Exposed to Urban Pollutants

Background: Urban air pollution contributes to lung and cardiovascular system dysfunction, making it a major concern for human health. Its impact on skin integrity, associated with increased occurrence of atopic dermatitis, is now recognized, but its cellular mechanisms remain poorly understood. Objective: In the present study we aimed at establishing the impact of urban pollutant on mitochondrial dynamics and bioenergetics using the HaCaT cell model. We also sought to establish the protective effect of ECH-5195 (red Panax ginseng extract), standardized in ginsenosides, in reversing pollution-induced mitochondrial defects. Methods: Urban pollution exposure was mimicked by 1 h exposure of HaCaT cells with standardized atmospheric particulate matter containing PAHs, nitro-PAHs, PCB congeners, and chlorinated pesticides with a mean particulate diameter of 5.85 μm (SRM1648). Results: The presence of urban pollutant in the cultures increased the prevalence of hyperfission by 1.41-fold (p = 0.023) and fission by 1.35 fold (p = 0.006) in the reticular mitochondrial network. ECH-5195 reduced both pollution-induced hyperfission by 0.54-fold (p = 0.004) and fission by 0.68-fold (p = 0.0006) normalizing the mitochondrial reticular network. Pollution exposure was associated with a significant reduction of basal OCR and increased lactate production, pushing the cell to rely on glycolysis for ATP production. When ECH-5195 was used, OCR was significantly increased, and the glycolytic contribution to ATP production was reduced while both oxidative phosphorylation and mitochondrial respiration were increased demonstrating mitochondrial re-engagement in ATP production. Conclusions: Pollution exposure was disruptive for both the mitochondrial network dynamics and mitochondrial respiration. Ginsenosides in ECH-5195 efficiently protected both from pollution-induced defects.

Abstract PO-026: Analysis of metabolic adaptations in HeLa cells as a function of energy status using a novel, direct ATP chelation methodology

Abstract Introduction: Since pioneering studies by Otto Warburg demonstrated that cancer cells had markedly reduced oxidative phosphorylation under conditions of high oxygen tension (Warburg Effect/aerobic glycolysis), metabolic reprogramming has become a well-recognized hallmark of cancer. While numerous discoveries have revealed highly specific metabolic differences between cancer cells, utilization of aerobic glycolysis as the primary means of ATP generation remains one of the most common metabolic alterations observed across a range of cancers. Gain-of-function mutations in p53 (mutp53) occur with a frequency of greater than 50% in many types of cancers and promote aerobic glycolysis by activation of glycolysis and inhibition of the tricarboxylic acid cycle. However, the manner by which these mutp53 regulates metabolic shifts in response to energy strain is only beginning to be elucidated. Hypothesis: Increased energy demand by cells caused by direct chelation of ATP by the novel, synthetic ATP-binding protein DX will be met by increases primarily in oxygen consumption. Methods: A cationic lipid mixture was complexed with active, purified DX protein to generate a DX/lipid complex. HeLa cells were incubated with the DX/lipid complex to allow efficient delivery of DX into the cytoplasm of the HeLa cells. The XF Real-Time ATP Rate Assay using Seahorse XF label-free technology was used to measure the rate of ATP production following DX expression and estimate the relative contributions of glycolysis and oxidative phosphorylation to the total ATP production rate. Results: HeLa cells transfected with DX had reduced viability, altered morphology, and decreased bioavailable intracellular ATP following DX expression. Unexpectedly, in response to DX expression and reduction in ATP, total ATP production rates in HeLa cells significantly decreased over time. Importantly, this drop in total ATP production was associated with a metabolic pathway utilization shift. Conclusion: The synthetically derived DX protein is a tool that allows for direct control of the ATP levels in cancer cells and has the potential to reveal novel metabolic adaptive responses to energy stress. Following regulated ATP reduction in HeLa cells, changes observed in the OXPHOS/glycolytic balance represent an overall cellular metabolic shift. Citation Format: Ashley P. Brown, Taha Muhammad, Selina Martinez, Shaleen B. Korch, Jeffrey Norris. Analysis of metabolic adaptations in HeLa cells as a function of energy status using a novel, direct ATP chelation methodology [abstract]. In: Abstracts: AACR Special Virtual Conference on Epigenetics and Metabolism; October 15-16, 2020; 2020 Oct 15-16. Philadelphia (PA): AACR; Cancer Res 2020;80(23 Suppl):Abstract nr PO-026.

Sodium Bicarbonate Ingestion Improves Time-to-Exhaustion Cycling Performance and Alters Estimated Energy System Contribution: A Dose-Response Investigation.

This study investigated the effects of two sodium bicarbonate (NaHCO3) doses on estimated energy system contribution and performance during an intermittent high-intensity cycling test (HICT), and time-to-exhaustion (TTE) exercise. Twelve healthy males (stature: 1.75 ± 0.08 m; body mass: 67.5 ± 6.3 kg; age: 21.0 ± 1.4 years; maximal oxygen consumption: 45.1 ± 7.0 ml.kg.min−1) attended four separate laboratory visits. Maximal aerobic power (MAP) was identified from an incremental exercise test. During the three experimental visits, participants ingested either 0.2 g.kg−1 BM NaHCO3 (SBC2), 0.3 g.kg−1 BM NaHCO3 (SBC3), or 0.07 g.kg−1 BM sodium chloride (placebo; PLA) at 60 min pre-exercise. The HICT involved 3 × 60 s cycling bouts (90, 95, 100% MAP) interspersed with 90 s recovery, followed by TTE cycling at 105% MAP. Blood lactate was measured after each cycling bout to calculate estimates for glycolytic contribution to exercise. Gastrointestinal (GI) upset was quantified at baseline, 30 and 60 min post-ingestion, and 5 min post-exercise. Cycling TTE increased for SBC2 (+20.2 s; p = 0.045) and SBC3 (+31.9 s; p = 0.004) compared to PLA. Glycolytic contribution increased, albeit non-significantly, during the TTE protocol for SBC2 (+7.77 kJ; p = 0.10) and SBC3 (+7.95 kJ; p = 0.07) compared to PLA. GI upset was exacerbated post-exercise after SBC3 for nausea compared to SBC2 and PLA (p < 0.05), whilst SBC2 was not significantly different to PLA for any symptom (p > 0.05). Both NaHCO3 doses enhanced cycling performance and glycolytic contribution, however, higher doses may maximize ergogenic benefits.

Multi-ingredient pre-workout supplementation changes energy system contribution and improves performance during high-intensity intermittent exercise in physically active individuals: a double-blind and placebo controlled study

BackgroundNutritional ergogenic aids are commonly used to boost physiological adaptations to exercise and promote greater fitness gains. However, there is a paucity of data about multi-ingredient pre-workout supplementation (MIPS). Therefore, the aim of the present study was to investigate the acute effects of MIPS on the oxidative, glycolytic and ATP-CP energy systems contribution, time spent above 90% V̇O2max (T90% V̇O2max), excess post-exercise oxygen consumption (EPOC) magnitude, number of efforts and time to exhaustion during a high-intensity interval exercise (HIIE) session.MethodsTwelve physically-active and healthy men completed the HIIE sessions that involved running bouts of 15 s on the treadmill at 120% of the maximum aerobic speed (MAS), interspersed with 15 s of passive recovery. Blood lactate was collected at immediately post, 3, 5, and 7 min post exercise. The contribution of ATP-CP, glycolytic and oxidative systems was analyzed at rest, during the HIIE sessions and for 20 min post. Performance variables (time to exhaustion, number of efforts) and oxygen consumption were also analyzed.ResultsMIPS significantly increased the number of efforts performed (MIPS: 41 ± 10 vs Placebo: 36 ± 12, p = 0.0220) and time to exhaustion (MIPS: 20.1 ± 6 min vs Placebo: 17 ± 5 min, p = 0.0226). There was no difference between supplements for both T90% V̇O2max (p = 0.9705) and EPOC (p = 0.4930). Consuming MIPS significantly increased the absolute oxidative energy system contribution by 23.8% (p = 0.0163) and the absolute ATP-CP contribution by 28.4% (p = 0.0055) compared to placebo. There was only a non-significant tendency for a higher glycolytic system contribution after MIPS ingestion (p = 0.0683).ConclusionAcute MIPS ingestion appears effective at increasing both aerobic and anaerobic alactic energy contribution and time to exhaustion during a HIIE protocol.

Chondroprotective effects of 4-methylumbelliferone and hyaluronan synthase-2 overexpression involve changes in chondrocyte energy metabolism

Hyaluronan is a critical component of articular cartilage and partially helps retain aggrecan within the extracellular matrix of this tissue. During osteoarthritis, hyaluronan and aggrecan loss are an early sign of tissue damage. However, our recent attempts to mimic hyaluronan loss with the hyaluronan inhibitor 4-methylumbelliferone (4MU) did not exacerbate arthritis-like features of in vitro models of arthritis, but surprisingly, caused the reverse (i.e. provided potent chondroprotection). Moreover, the protective effects of 4MU did not depend on its role as a hyaluronan inhibitor. To understand the molecular mechanism in 4MU-mediated chondroprotection, we considered recent studies suggesting that shifts in intracellular UDP-hexose pools promote changes in metabolism. To determine whether such metabolic shifts are associated with the mechanism of 4MU-mediated pro-catabolic inhibition, using molecular and metabolomics approaches, we examined whether bovine and human chondrocytes exhibit changes in the contribution of glycolysis and mitochondrial respiration to ATP production rates as well as in other factors that respond to or might drive these changes. Overexpression of either HA synthase-2 or 4MU effectively reduced dependence on glycolysis in chondrocytes, especially enhancing glycolysis use by interleukin-1β (IL1β)-activated chondrocytes. The reduction in glycolysis secondarily enhanced mitochondrial respiration in chondrocytes, which, in turn, rescued phospho-AMP-activated protein kinase (AMPK) levels in the activated chondrocytes. Other glycolysis inhibitors, unrelated to hyaluronan biosynthesis, namely 2-deoxyglucose and dichloroacetate, caused metabolic changes in chondrocytes equivalent to those elicited by 4MU and similarly protected both chondrocytes and cartilage explants. These results suggest that fluxes in UDP-hexoses alter metabolic energy pathways in cartilage.