Zbigniew Szkutnik

Training‐induced acceleration of O2 uptake on‐kinetics precedes muscle mitochondrial biogenesis in humans

•  What is the central question of this study? A few weeks of endurance training accelerate the oxygen uptake () on‐kinetics in humans. The main aim of the present study was to determine whether the acceleration of on‐kinetics obtained by a short period of moderate‐intensity training can be explained by an intensification of mitochondrial biogenesis. •  What is the main finding and its importance? We demonstrated that 5 weeks of moderate‐intensity training accelerates the on‐kinetics during moderate‐intensity cycling in the absence of enhanced mitochondrial biogenesis or capillarization in the trained muscles. We postulate that in the early stages of training an intensification of ‘parallel activation’ of oxidative phosphorylation could account for the shortening of the on‐transient.

Detection of the change point in oxygen uptake during an incremental exercise test using recursive residuals: relationship to the plasma lactate accumulation and blood acid base balance

Abstract The purpose of this study was to develop a method to determine the power output at which oxygen uptake (V˙O2) during an incremental exercise test begins to rise non-linearly. A group of 26 healthy non-smoking men [mean age 22.1 (SD 1.4) years, body mass 73.6 (SD 7.4) kg, height 179.4 (SD 7.5) cm, maximal oxygen uptake (V˙O2max) 3.726 (SD 0.363) l · min−1], experienced in laboratory tests, were the subjects in this study. They performed an incremental exercise test on a cycle ergometer at a pedalling rate of 70 rev · min−1. The test started at a power output of 30 W, followed by increases amounting to 30 W every 3 min. At 5 min prior to the first exercise intensity, at the end of each stage of exercise protocol, blood samples (1 ml each) were taken from an antecubital vein. The samples were analysed for plasma lactate concentration [La]pl, partial pressure of O2 and CO2 and hydrogen ion concentration [H+]b. The lactate threshold (LT) in this study was defined as the highest power output above which [La−]pl showed a sustained increase of more than 0.5 mmol · l−1 · step−1. The V˙O2 was measured breath-by-breath. In the analysis of the change point (CP) of V˙O2 during the incremental exercise test, a two-phase model was assumed for the 3rd-min-data of each step of the test: Xi=ati+b+ɛi for i=1,2,…,T, and E(Xi)>ati+b for i =T+1,…,n, where X1, … , Xn are independent and ɛi∼N(0,σ2). In the first phase, a linear relationship between V˙O2 and power output was assumed, whereas in the second phase an additional increase in V˙O2 above the values expected from the linear model was allowed. The power output at which the first phase ended was called the change point in oxygen uptake (CP-V˙O2). The identification of the model consisted of two steps: testing for the existence of CP and estimating its location. Both procedures were based on suitably normalised recursive residuals. We showed that in 25 out of 26 subjects it was possible to determine the CP-O2 as described in our model. The power output at CP-V˙O2 amounted to 136.8 (SD 31.3) W. It was only 11 W – non significantly – higher than the power output corresponding to LT. The V˙O2 at CP-V˙O2 amounted to 1.828 (SD 0.356) l · min−1 was [48.9 (SD 7.9)% V˙O2max]. The [La−]pl at CP-V˙O2, amounting to 2.57 (SD 0.69) mmol · l−1 was significantly elevated (P<0.01) above the resting level [1.85 (SD 0.46) mmol · l−1], however the [H+]b at CP-V˙O2 amounting to 45.1 (SD 3.0) nmol · l−1, was not significantly different from the values at rest which amounted to 44.14 (SD 2.79) nmol · l−1. An increase of power output of 30 W above CP-V˙O2 was accompanied by a significant increase in [H+]b above the resting level (P=0.03).

Mechanisms responsible for the acceleration of pulmonary V̇o2 on-kinetics in humans after prolonged endurance training

The effect of prolonged endurance training on the pulmonary V̇O2 on- and off-kinetics in humans, in relation to muscle mitochondria biogenesis, is investigated. Eleven untrained physically active men (means±SD: age 22.4±1.5 years, V̇O2peak 3,187±479 ml/min) performed endurance cycling training (4 sessions per week) lasting 20 wk. Training shortened τp of the pulmonary V̇O2 on-kinetics during moderate-intensity cycling by ∼19% from 28.3±5.2 to 23.0±4.0 s (P=0.005). τp of the pulmonary V̇O2 off-kinetics decreased by ∼11% from 33.7±7.2 to 30.0±6.6 (P=0.02). Training increased (in vastus lateralis muscle) mitochondrial DNA copy number in relation to nuclear DNA (mtDNA/nDNA) (+53%) (P=0.014), maximal citrate synthase (CS) activity (+38%), and CS protein content (+38%) (P=0.004), whereas maximal cytochrome c oxidase (COX) activity after training tended to be only slightly (+5%) elevated (P=0.08). By applying to the experimental data, our computer model of oxidative phosphorylation (OXPHOS) and using metabolic control analysis, we argue that COX activity is a much better measure of OXPHOS intensity than CS activity. According to the model, in the present study a training-induced increase in OXPHOS activity accounted for about 0-10% of the decrease in τp of muscle and pulmonary V̇O2 for the on-transient, whereas the remaining 90-100% is caused by an increase in each-step parallel activation of OXPHOS.

Endurance training decreases the non-linearity in the oxygen uptake–power output relationship in humans

In this study, we hypothesized that 5 weeks of cycling endurance training can decrease the magnitude of the non‐proportional increase in oxygen uptake ( ) to power output relationship ( ‘excess’) at exercise intensities exceeding the lactate threshold (LT). Ten untrained, physically active men performed a bout of incremental cycling exercise until exhaustion before and after training. The mitochondrial DNA copy number, myosin heavy chain composition and content of uncoupling protein 3 and sarcoplasmic reticulum Ca2+‐ATPases (SERCAs) were analysed in muscle biopsies taken from vastus lateralis before and after training. The training resulted in an enhancement of the power‐generating capabilities at maximal oxygen uptake ( ) by ∼7% (P= 0.002) despite there being no changes in (P= 0.49). This effect was due to a considerable reduction in the magnitude of the ‘excess’ (P < 0.05) above the LT. A decrease in plasma ammonia concentration was found during exercise after training (P < 0.05). A downregulation of SERCA2 in vastus lateralis (P= 0.006) was observed after training. No changes in myosin heavy chain composition, selected electron transport chain proteins, uncoupling protein 3 or the mitochondrial DNA copy number (P > 0.05) were found after training. We conclude that the training‐induced increase in power‐generating capabilities at was due to attenuation of the ‘excess’ above the LT. This adaptive response seems to be related to the improvement of muscle metabolic stability, as judged by a lowering of plasma ammonia concentration. The enhancement of muscle metabolic stability after training could be caused by a decrease in ATP usage at a given power output owing to downregulation of SERCA2 pumps.

Mechanisms of Attenuation of Pulmonary V'O2 Slow Component in Humans after Prolonged Endurance Training.

In this study we have examined the effect of prolonged endurance training program on the pulmonary oxygen uptake (V’O2) kinetics during heavy-intensity cycling-exercise and its impact on maximal cycling and running performance. Twelve healthy, physically active men (mean±SD: age 22.33±1.44 years, V’O2peak 3198±458 mL ∙ min-1) performed an endurance training composed mainly of moderate-intensity cycling, lasting 20 weeks. Training resulted in a decrease (by ~5%, P = 0.027) in V’O2 during prior low-intensity exercise (20 W) and in shortening of τp of the V’O2 on-kinetics (30.1±5.9 s vs. 25.4±1.5 s, P = 0.007) during subsequent heavy-intensity cycling. This was accompanied by a decrease of the slow component of V’O2 on-kinetics by 49% (P = 0.001) and a decrease in the end-exercise V’O2 by ~5% (P = 0.005). An increase (P = 0.02) in the vascular endothelial growth factor receptor 2 mRNA level and a tendency (P = 0.06) to higher capillary-to-fiber ratio in the vastus lateralis muscle were found after training (n = 11). No significant effect of training on the V’O2peak was found (P = 0.12). However, the power output reached at the lactate threshold increased by 19% (P = 0.01). The power output obtained at the V’O2peak increased by 14% (P = 0.003) and the time of 1,500-m performance decreased by 5% (P = 0.001). Computer modeling of the skeletal muscle bioenergetic system suggests that the training-induced decrease in the slow component of V’O2 on-kinetics found in the present study is mainly caused by two factors: an intensification of the each-step activation (ESA) of oxidative phosphorylation (OXPHOS) complexes after training and decrease in the ‘‘additional” ATP usage rising gradually during heavy-intensity exercise.

Change-point detection in a shape-restricted regression model

A regression model with a possible structural change and with a small number of measurements is considered. A priori information about the shape of the regression function is used to formulate the model as a linear regression model with inequality constraints and a likelihood ratio test for the presence of a change-point is constructed. The exact null distribution of the test statistic is given. Consistency of the test is proved when the noise level goes to zero. Numerical approximations to the powers against various alternatives are given and compared with the powers of the k-linear-r-ahead recursive residuals tests and CUSUM tests. Performance of four different estimators of the change-point is studied in a Monte Carlo experiment. An application of the procedures to some real data is also presented.

On the Durbin-Wagle randomization device and some of its applications

Validity of Wagles multivariate extension of the Durbin randomization device is directly proved and some vague points of the original paper are clarified. The device is then used in a non-standard way to obtain asymptotic distributions of some functions of the multivariate Gaussian sample configuration. Applications comprise, e.g., null distributions of some statistics naturally emerging in the context of invariant testing multivariate normality.

Adaptation of motor unit contractile properties in rat medial gastrocnemius to treadmill endurance training: Relationship to muscle mitochondrial biogenesis

This study aimed at investigating the effects of 2, 4 and 8 weeks of endurance training on the contractile properties of slow (S), fast fatigue resistant (FR) and fast fatigable (FF) motor units (MUs) in rat medial gastrocnemius (MG) in relation to the changes in muscle mitochondrial biogenesis. The properties of functionally isolated MUs were examined in vivo. Mitochondrial biogenesis was judged based on the changes in mitochondrial DNA copy number (mtDNA), the content of the electron transport chain (ETC) proteins and PGC-1α in the MG. Moreover, the markers of mitochondria remodeling mitofusins (Mfn1, Mfn2) and dynamin-like protein (Opa1) were studied using qPCR. A proportion of FR MUs increased from 37.9% to 50.8% and a proportion of FF units decreased from 44.7% to 26.6% after 8 weeks of training. The increased fatigue resistance, shortened twitch duration, and increased ability to potentiate force were found as early as after 2 weeks of endurance training, predominantly in FR MUs. Moreover, just after 2 weeks of the training an enhancement of the mitochondrial network remodeling was present as judged by an increase in expression of Mfn1, Opa1 and an increase in PGC-1α in the slow part of MG. Interestingly, no signs of intensification of mitochondrial biogenesis assessed by ETC proteins content and mtDNA in slow and fast parts of gastrocnemius were found at this stage of the training. Nevertheless, after 8 weeks of training an increase in the ETC protein content was observed, but mainly in the slow part of gastrocnemius. Concluding, the functional changes in MUs’ contractile properties leading to the enhancement of muscle performance accompanied by an activation of signalling that controls the muscle mitochondrial network reorganisation and mitochondrial biogenesis belong to an early muscle adaptive responses that precede an increase in mitochondrial ETC protein content.

Effect of temperature on fatty acid metabolism in skeletal muscle mitochondria of untrained and endurance-trained rats

We studied the effects of various assay temperatures, representing hypothermia (25°C), normothermia (35°C), and hyperthermia (42°C), on the oxidation of lipid-derived fuels in rat skeletal muscle mitochondria of untrained and endurance-trained rats. Adult 4-month-old male Wistar rats were assigned to a training group (rats trained on a treadmill for 8 weeks) or a sedentary control group. In skeletal muscle mitochondria of both control and trained rats, an increase in the assay temperature from 25°C to 42°C was accompanied by a consistent increase in the oxidation of palmitoylcarnitine and glycerol-3-phosphate. Moreover, endurance training increased mitochondrial capacity to oxidize the lipid-derived fuels at all studied temperatures. The endurance training-induced increase in mitochondrial capacity to oxidize fatty acids was accompanied by an enhancement of mitochondrial biogenesis, as shown by the elevated expression levels of Nrf2, PGC1α, and mitochondrial marker and by the elevated expression levels of mitochondrial proteins involved in fatty acid metabolism, such as fatty acid transporter CD36, carnitine palmitoyltransferase 1A (CPT1A), and acyl-CoA dehydrogenase (ACADS). We conclude that hyperthermia enhances but hypothermia attenuates the rate of the oxidation of fatty acids and glycerol-3-phosphate in rat skeletal muscle mitochondria isolated from both untrained and trained rats. Moreover, our results indicate that endurance training up-regulates mitochondrial biogenesis markers, lipid-sustained oxidative capacity, and CD36 and CPT1A proteins involved in fatty acid transport, possibly via PGC1α and Nrf2 signaling pathways.