L. Bruce Gladden

Oxygen uptake on-kinetics in dog gastrocnemius in situ following activation of pyruvate dehydrogenase by dichloroacetate.

The aim of the present study was to determine whether the activation of the pyruvate dehydrogenase complex (PDC) by dichloroacetate (DCA) is associated with faster O2 uptake (V̇O2) on‐kinetics. V̇O2 on‐kinetics was determined in isolated canine gastrocnemius muscles in situ (n= 6) during the transition from rest to 4 min of electrically stimulated isometric tetanic contractions, corresponding to ∼60–70 % of peak V̇O2. Two conditions were compared: (1) control (saline infusion, C); and (2) DCA infusion (300 mg (kg body mass)−1, 45 min before contraction). Muscle blood flow (Q̇) was measured continuously in the popliteal vein; arterial and popliteal vein O2 contents were measured at rest and at 5–7 s intervals during the transition. Muscle V̇O2 was calculated as Q̇ multiplied by the arteriovenous O2 content difference. Muscle biopsies were taken before and at the end of contraction for determination of muscle metabolite concentrations. DCA activated PDC at rest, as shown by the 9‐fold higher acetylcarnitine concentration in DCA (vs. C; P < 0.0001). Phosphocreatine degradation and muscle lactate accumulation were not significantly different between C and DCA. DCA was associated with significantly less muscle fatigue. Resting and steady‐state V̇O2 values during contraction were not significantly different between C and DCA. The time to reach 63 % of the V̇O2 difference between the resting baseline and the steady‐state V̇O2 values during contraction was 22.3 ± 0.5 s in C and 24.5 ± 1.4 s in DCA (n.s.). In this experimental model, activation of PDC by DCA resulted in a stockpiling of acetyl groups at rest and less muscle fatigue, but it did not affect ‘anaerobic’ energy provision and V̇O2 on‐kinetics.

Blood Lactate Measurements and Analysis during Exercise: A Guide for Clinicians

Blood lactate concentration ([La−]b) is one of the most often measured parameters during clinical exercise testing as well as during performance testing of athletes. While an elevated [Lai may be indicative of ischemia or hypoxemia, it may also be a “normal” physiological response to exertion. In response to “all-out” maximal exertion lasting 30–120 seconds, peak [La−]b values of =15–25 mM may be observed 3–8 minutes postexercise. In response to progressive, incremental exercise, [La−]b increases gradually at first and then more rapidly as the exercise becomes more intense. The work rate beyond which [La−]b increases exponentially [the lactate threshold (LT)] is a better predictor of performance than VO2max and is a better indicator of exercise intensity than heart rate; thus LT (and other valid methods of describing this curvilinear [La−]b response with a single point) is useful in prescribing exercise intensities for most diseased and nondiseased patients alike. H+-monocarboxylate cotransporters provide the primary of three routes by which La− transport proceeds across the sarcolemma and red blood cell membrane. At rest and during most exercise conditions, whole blood [La−] values are on average 70% of the corresponding plasma [La−] values; thus when analyzing [La−]b, care should be taken to both (1) validate the [LaT-measuring instrument with the criterion/reference enzymatic method and (2) interpret the results correctly based on what is being measured (plasma or whole blood). Overall, it is advantageous for clinicians to have a thorough understanding of [La−] responses, blood La− transport and distribution, and [La−] analysis.

Comparison of the BOD POD with the four-compartment model in adult females.

PURPOSE This study was designed to compare the accuracy and bias in estimates of total body density (Db) by hydrostatic weighing (HW) and the BOD POD, and percent body fat (%fat) by the BOD POD with the four-compartment model (4C model) in 42 adult females. Furthermore, the role of the aqueous and mineral fractions in the estimation of body fat by the BOD POD was examined. METHODS Total body water was determined by isotope dilution ((2)H(2)0) and bone mineral was determined by dual-energy x-ray absorptiometry. Db and %fat were determined by the BOD POD and HW. The 4C model of Baumgartner was used as the criterion measure of body fat. RESULTS HW Db (1.0352 g x cm(-3)) was not statistically different (P = 0.35) from BOD POD Db (1.0349 g x cm(-3)). The regression between Db by HW and the BOD POD significantly deviated from the line of identity (Db by HW = 0.90 x Db by BOD POD + 0.099; R(2) = 0.94). BOD POD %fat (28.8%) was significantly lower (P < 0.01) than %fat by the 4C model (30.6%). The regression between %fat by the 4C model and the BOD POD significantly deviated from the line of identity (%fat by 4C model = 0.88 x %fat by BOD POD + 5.41%; R(2) = 0.92). BOD POD Db and %fat showed no bias across the range of fatness. Only the aqueous fraction of the fat-free mass (FFM) had a significant correlation with the difference in %fat between the 4C model and the BOD POD. CONCLUSION These data indicate that the BOD POD underpredicted body fat as compared with the 4C model, and the aqueous fraction of the FFM had a significant effect on estimates of %fat by the BOD POD.

Effects of nitric oxide synthase inhibition by l-NAME on oxygen uptake kinetics in isolated canine muscle in situ

Nitric oxide (NO) has an inhibitory action on O2 uptake at the level of the mitochondrial respiratory chain. The aim of this study was to evaluate the effects of NO synthase (NOS) inhibition on muscle kinetics. Isolated canine gastrocnemius muscles in situ (n= 6) were studied during transitions from rest to 4‐min of electrically stimulated contractions corresponding to ∼60% of the muscle peak . Two conditions were compared: (i) Control (CTRL) and (ii) l‐NAME, in which the NOS inhibitor l‐NAME (20 mg kg−1) was administered. In both conditions the muscle was pump‐perfused with constantly elevated blood flow , at a level measured during a preliminary contraction trial with spontaneous self‐perfused. A vasodilatory drug was also infused. Arterial and venous O2 concentrations were determined at rest and at 5–7 s intervals during the transition. was calculated by Ficks principle. Muscle biopsies were obtained at rest and during contractions. Muscle force was measured continuously. Phosphocreatine hydrolysis and the calculated substrate level phosphorylation were slightly (but not significantly) lower in l‐NAME than in CTRL. Significantly (P < 0.05) less fatigue was found in l‐NAME versus CTRL. The time delay (TDf) and the time constant (τf) of the ‘fundamental’ component of kinetics were not significantly different between CTRL (TDf 7.2 ± 1.2 s; and τf 10.6 ± 1.3, ±s.e.m.) and l‐NAME (TDf 9.3 ± 0.6; and τf 10.4 ± 1.0). Contrary to our hypothesis, NOS inhibition did not accelerate muscle kinetics. The down‐regulation of mitochondrial respiration by NO does not limit the kinetics of adjustment of oxidative metabolism at exercise onset.

Muscle Glycogen Resynthesis after Short Term, High Intensity Exercise and Resistance Exercise

SummaryTypical rates of muscle glycogen resynthesis after short term, high intensity exercise (15.1 to 33.6 mmol/kg/h) are much higher than glycogen resynthesis rates following prolonged exercise (≈2 mmol/kg/h), even when optimal amounts of oral carbohydrate are supplied (≈8 mmol/kg/h). Several factors differ during post-exercise recovery from short term, high intensity exercise compared with prolonged exercise. The extremely fast rate of muscle glycogen resynthesis following short term, high intensity exercise may originate from these differences.First, peak blood glucose levels range from 6.6 to 8.9 mmol/L during recovery from short term, high intensity exercise. This is markedly higher than the blood glucose values of 2 to 3.4 mmol/L after prolonged exercise. In response to this elevation in plasma glucose levels, insulin levels increase to ≈60 μU/ml, a 2-fold increase over resting values. Both glucose and insulin regulate glycogen synthase activity, and higher levels of them improve muscle glycogen synthesis. Secondly, high intensity exercise produces high levels of glycolytic intermediates in muscle, as well as high lactate levels ([La]) in muscle and blood. Finally, fast-twitch glycolytic muscle fibres are more heavily used in short term, high intensity exercise. This promotes greater glycogen depletion in the fast-twitch fibres, which have a higher level of glycogen synthase activity than slow-twitch fibres.While the exact contribution of each of these factors is unknown, they may act in combination to stimulate rapid muscle glycogen resynthesis rates. Muscle glycogen resynthesis rates following resistance exercise (1.3 to 11.1 mmol/kg/h) are slower than the rates observed after short term, high intensity exercise. This may be caused by slightly lower muscle and blood [La] after resistance exercise. In addition, a greater eccentric component in the resistance exercise may cause some interference with glycogen resynthesis.

Kinetic control of oxygen consumption during contractions in self-perfused skeletal muscle

Non‐technical summary  The ability to sustain exercise is dependent on the ability to match muscular energy supply fuelled by oxygen to the energy demands of the activity (the ‘currency’ of biological energy is termed ATP). Experiments using muscle samples in a test tube suggest that the activation of muscle oxygen consumption is caused by accumulation of ADP (a breakdown product of ATP), which signals the need to increase ATP supply. The mechanism of this signalling process in vivo, however, is not well understood. We investigated the mechanism controlling oxidative ATP supply activation in canine muscle, by simultaneous measurements of oxygen consumption and ADP at the onset of muscle contractions. At the start of contractions muscle oxygen consumption increased more rapidly than predicted from the measured accumulation of ADP. These data suggest that an additional process (or processes) occurs to activate muscle oxidative ATP provision at the onset of exercise in vivo.

Lactate is always the end product of glycolysis

Through much of the history of metabolism, lactate (La−) has been considered merely a dead-end waste product during periods of dysoxia. Congruently, the end product of glycolysis has been viewed dichotomously: pyruvate in the presence of adequate oxygenation, La− in the absence of adequate oxygenation. In contrast, given the near-equilibrium nature of the lactate dehydrogenase (LDH) reaction and that LDH has a much higher activity than the putative regulatory enzymes of the glycolytic and oxidative pathways, we contend that La− is always the end product of glycolysis. Cellular La− accumulation, as opposed to flux, is dependent on (1) the rate of glycolysis, (2) oxidative enzyme activity, (3) cellular O2 level, and (4) the net rate of La− transport into (influx) or out of (efflux) the cell. For intracellular metabolism, we reintroduce the Cytosol-to-Mitochondria Lactate Shuttle. Our proposition, analogous to the phosphocreatine shuttle, purports that pyruvate, NAD+, NADH, and La− are held uniformly near equilibrium throughout the cell cytosol due to the high activity of LDH. La− is always the end product of glycolysis and represents the primary diffusing species capable of spatially linking glycolysis to oxidative phosphorylation.

Modeling oxygenation in venous blood and skeletal muscle in response to exercise using near-infrared spectroscopy

Noninvasive, continuous measurements in vivo are commonly used to make inferences about mechanisms controlling internal and external respiration during exercise. In particular, the dynamic response of muscle oxygenation (Sm(O(2))) measured by near-infrared spectroscopy (NIRS) is assumed to be correlated to that of venous oxygen saturation (Sv(O(2))) measured invasively. However, there are situations where the dynamics of Sm(O(2)) and Sv(O(2)) do not follow the same pattern. A quantitative analysis of venous and muscle oxygenation dynamics during exercise is necessary to explain the links between different patterns observed experimentally. For this purpose, a mathematical model of oxygen transport and utilization that accounts for the relative contribution of hemoglobin (Hb) and myoglobin (Mb) to the NIRS signal was developed. This model includes changes in microvascular composition within skeletal muscle during exercise and integrates experimental data in a consistent and mechanistic manner. Three subjects (age 25.6 +/- 0.6 yr) performed square-wave moderate exercise on a cycle ergometer under normoxic and hypoxic conditions while muscle oxygenation (C(oxy)) and deoxygenation (C(deoxy)) were measured by NIRS. Under normoxia, the oxygenated Hb/Mb concentration (C(oxy)) drops rapidly at the onset of exercise and then increases monotonically. Under hypoxia, C(oxy) decreases exponentially to a steady state within approximately 2 min. In contrast, model simulations of venous oxygen concentration show an exponential decrease under both conditions due to the imbalance between oxygen delivery and consumption at the onset of exercise. Also, model simulations that distinguish the dynamic responses of oxy-and deoxygenated Hb (HbO(2), HHb) and Mb (MbO(2), HMb) concentrations (C(oxy) = HbO(2) + MbO(2); C(deoxy) = HHb + HMb) show that Hb and Mb contributions to the NIRS signal are comparable. Analysis of NIRS signal components during exercise with a mechanistic model of oxygen transport and metabolism indicates that changes in oxygenated Hb and Mb are responsible for different patterns of Sm(O(2)) and Sv(O(2)) dynamics observed under normoxia and hypoxia.

Faster O2 uptake kinetics in canine skeletal muscle in situ after acute creatine kinase inhibition

The ability to sustain skeletal muscle contractions is dependent on the conversion of chemical to mechanical energy – a process fueled by adenosine triphosphate (ATP). The link between two of the major mechanisms for ATP provision, phosphocreatine (PCr) breakdown and oxidative phosphorylation, was investigated in canine muscle. Infusion of a drug to prevent PCr breakdown (via inhibition of the enzyme creatine kinase; CK) caused (among other effects) a faster adjustment of energy provision from oxidation upon the onset of contractions. Thus, in mammalian skeletal muscle the CK enzyme slows the signal responsible for the activation of oxidative phosphorylation. Sudden increases in the demands for energy at the onset of exercise are met by PCr breakdown, but this process is functionally related, presumably through the levels of some of its metabolites, to the regulation of oxidative phosphorylation, the most important pathway for ATP resynthesis.

A prior bout of contractions speeds V̇o2 and blood flow on-kinetics and reduces the V̇o2 slow-component amplitude in canine skeletal muscle contracting in situ

It was the purpose of this study to examine the effect of a priming contractile bout on oxygen uptake (VO2) on-kinetics in highly oxidative skeletal muscle. Canine gastrocnemii (n=12) were stimulated via their sciatic nerves (8 V, 0.2-ms duration, 50 Hz, 200-ms train) at a rate of 2 contractions/3 s (approximately 70% peak VO2) for two 2-min bouts, separated by 2 min of recovery. Blood flow was recorded with an ultrasonic flowmeter, and muscle oxygenation monitored via near-infrared spectroscopy. Compared with the first bout (bout 2 vs. bout 1), the VO2 primary time constant (mean+/-SD, 9.4+/-2.3 vs. 12.0+/-3.9 s) and slow-component amplitude (5.9+/-6.3 vs. 12.1+/-9.0 ml O2.kg wet wt(-1).min(-1)) were significantly reduced (P<0.05) during the second bout. Blood flow on-kinetics were significantly speeded during the second bout (time constant=7.7+/-2.6 vs. 14.8+/-5.8 s), and O2 extraction was greater at the onset of contractions (0.050+/-0.030 vs. 0.020+/-0.010 ml O2/ml blood). Kinetics of muscle deoxygenation were significantly slower at the onset of the second bout (7.2+/-2.2 vs. 4.4+/-1.2 s), while relative oxyhemoglobin concentration was elevated throughout the second bout. These results suggest that better matching of O2 delivery to VO2 speeds Vo(2) on-kinetics at this metabolic rate, but do not eliminate a potential role for enhanced metabolic activation. Additionally, altered motor unit recruitment at the onset of a second bout is not a prerequisite for reductions in the VO2 slow-component amplitude after a priming contractile bout in canine muscle in situ.