Andreas Nieß

Short-Term Treadmill Running as a Model for Studying Cell-Free DNA Kinetics In Vivo

BACKGROUND Increased plasma concentrations of cell-free DNA (cf-DNA) are considered a hallmark of various clinical conditions. Despite intensive research in this field, limited data are available concerning the time course of release and clearance of cf-DNA in vivo. METHODS We extracted cf-DNA from plasma samples taken before and immediately after a 10-km cross-country run, and from samples taken before, immediately after, and 30 min after exhaustive short-term treadmill exercise. The contribution of nuclear (nDNA) and mitochondrial DNA (mtDNA) was measured by quantitative real-time PCR. The incremental treadmill exercise setup was exploited to delineate the precise sequencing and timing of cf-nDNA, lactate, and high-mobility group box 1 protein (HMGB1) release during the exercise and recovery phases. RESULTS Postexercise plasma cf-nDNA concentrations in cross-country and treadmill runners were significantly increased, by 7.6-fold and 9.9-fold, respectively (P < 0.001). cf-nDNA concentrations were not correlated with age, sex, or body mass index. Plasma concentrations of cf-nDNA and HMGB1 in postexercise samples of treadmill runners were significantly correlated (r = 0.84; P = 0.004). cf-mtDNA concentrations were not affected by treadmill exercise. Time-course analyses demonstrated that cf-nDNA is released within minutes after the onset of exercise and is rapidly cleared from the circulation after the cessation of exercise. Nearly congruent kinetics for cf-nDNA, lactate, and HMGB1 were observed during the exercise phase. CONCLUSIONS A single bout of exhaustive short-term treadmill exercise constitutes a versatile model system suitable for addressing basic questions about cf-DNA biology.

Basal and exercise induced label-free quantitative protein profiling of m. vastus lateralis in trained and untrained individuals.

UNLABELLED Morphological and metabolic adaptations of the human skeletal muscle to exercise are crucial to improve performance and prevent chronic diseases and metabolic disorders. In this study we investigated human skeletal muscle protein composition in endurance trained (ET) versus untrained individuals (UT) and its modulation by an acute bout of endurance exercise. Participants were recruited based on their VO2max and subjected to a bicycle exercise test. M. vastus lateralis biopsies were taken before and three hours after exercise. Muscle lysates were analyzed using off-gel LC-MS/MS. Relative protein abundances were compared between ET and UT at rest and after exercise. Comparing UT and ET, we identified 92 significantly changed proteins under resting conditions. Specifically, fiber-type-specific and proteins of the oxidative phosphorylation and tricarboxylic acid cycle were increased in ET. In response to acute exercise, 71 proteins in ET and 44 in UT were altered. Here, a decrease of proteins involved in energy metabolism accompanied with alterations of heat shock and proteasomal proteins could be observed. In summary, long-term endurance training increased the basal level of structural and mitochondrial proteins in skeletal muscle. In contrast, acute exercise resulted in a depletion of proteins related to substrate utilization, especially in trained athletes. BIOLOGICAL SIGNIFICANCE The investigation of the human skeletal muscle proteome in response to exercise may provide novel insights into the process of muscular plasticity. It is of importance in the development of exercise-based strategies in the prevention and therapy of many chronic inflammatory and degenerative diseases which are often accompanied by muscular deconditioning. Up to date, proteomic investigations of the human muscle proteome in adaptation to exercise are mainly focused on untrained individuals and often restricted to animal studies. In the present study we compare the protein composition in endurance trained athletes and untrained individuals in the resting muscle and its modulation in response to acute exercise. To our knowledge, we present the first comprehensive analysis of skeletal muscle proteome alterations in response to acute and long-term exercise intervention.

TGF-β Contributes to Impaired Exercise Response by Suppression of Mitochondrial Key Regulators in Skeletal Muscle.

A substantial number of people at risk of developing type 2 diabetes could not improve insulin sensitivity by physical training intervention. We studied the mechanisms of this impaired exercise response in 20 middle-aged individuals at high risk of developing type 2 diabetes who performed 8 weeks of controlled cycling and walking training at 80% individual Vo2 peak. Participants identified as nonresponders in insulin sensitivity (based on the Matsuda index) did not differ in preintervention parameters compared with high responders. The failure to increase insulin sensitivity after training correlates with impaired upregulation of mitochondrial fuel oxidation genes in skeletal muscle, and with the suppression of the upstream regulators PGC1α and AMPKα2. The muscle transcriptomes of the nonresponders are further characterized by the activation of transforming growth factor (TGF)-β and TGF-β target genes, which is associated with increases in inflammatory and macrophage markers. TGF-β1 as inhibitor of mitochondrial regulators and insulin signaling is validated in human skeletal muscle cells. Activated TGF-β1 signaling downregulates the abundance of PGC1α, AMPKα2, the mitochondrial transcription factor TFAM, and mitochondrial enzymes. Thus, the data suggest that increased TGF-β activity in skeletal muscle can attenuate the improvement of mitochondrial fuel oxidation after training and contribute to the failure to increase insulin sensitivity.

Heart rate variability biofeedback therapy and graded exercise training in management of chronic fatigue syndrome: An exploratory pilot study

OBJECTIVE Chronic fatigue syndrome (CFS) is characterised by persistent fatigue, exhaustion, and several physical complaints. Research has shown cognitive behavioural therapy (CBT) and graded exercise training (GET) to be the most effective treatments. In a first step we aimed to assess the efficacy of heart rate variability biofeedback therapy (HRV-BF) as a treatment method comprising cognitive and behavioural strategies and GET in the pilot trial. In a second step we aimed to compare both interventions with regard to specific parameters. METHODS The study was conducted in an outpatient treatment setting. A total of 28 women with CFS (50.3±9.3years) were randomly assigned to receive either eight sessions of HRV-BF or GET. The primary outcome was fatigue severity. Secondary outcomes were mental and physical quality of life and depression. Data were collected before and after the intervention as well as at a 5-month follow-up. RESULTS General fatigue improved significantly after both HRV-BF and GET. Specific cognitive components of fatigue, mental quality of life, and depression improved significantly after HRV-BF only. Physical quality of life improved significantly after GET. There were significant differences between groups regarding mental quality of life and depression favouring HRV-BF. CONCLUSION Both interventions reduce fatigue. HRV-BF seems to have additional effects on components of mental health, including depression, whereas GET seems to emphasise components of physical health. These data offer implications for further research on combining HRV-BF and GET in patients with CFS. TRIAL REGISTRATION The described trial has been registered at the International Clinical Trials Registry Platform following the number DRKS00005445.

Neutrophil Extracellular Traps: A Walk on the Wild Side of Exercise Immunology

Intense exercise evokes a rapid and transient increase in circulating cell-free DNA (cf-DNA), a phenomenon that is commonly observed in a variety of acute and chronic inflammatory conditions. While the potential value of cf-DNA for the prediction of disease outcome and therapeutic response is well documented, the release mechanisms and biological relevance of cf-DNA have long remained enigmatic. The discovery of neutrophil extracellular traps (NETs) provided a novel mechanistic explanation for increased cf-DNA levels. Now there is increasing evidence that NETs may contribute to cf-DNA in diverse infectious, non-infectious and autoinflammatory conditions, as well as in response to acute exercise. NETs have now been firmly established as a fundamental immune mechanism used by neutrophils to respond to infection and tissue injury. On the other side, aberrant formation of NETs appears to be a driving force in the pathogenesis of autoimmunity and cardiovascular disease. Thus, the emergence of NETs in the ‘exercising vasculature’ raises important questions considering beneficial effects, as well as occasional adverse effects, of exercise on immune homeostasis. This review gives an overview of the current state of research into the mechanisms of how NETs are released, contribute to host defence and participate in inflammatory disorders. We discuss the impact of exercise-induced NETs, considering a potentially beneficial role in the prevention of lifestyle-related diseases, as well as putative detrimental effects that may arise in elite sports. Finally, we propose that exercise-induced cf-DNA responses could be exploited for diagnostic/prognostic purposes to identify individuals who are at increased risk of cardiovascular events or autoimmunity.

Effects of extracellular orotic acid on acute contraction-induced adaptation patterns in C2C12 cells

Dietary administration of orotic acid (OA), an intermediate in the pyrimidine biosynthetic pathway, is considered to provide a wide range of beneficial effects, including cardioprotection and exercise adaptation. Its mechanisms of action, when applied extracellularly, however, are barely understood. In this study, we evaluated potential effects of OA on skeletal muscle using an in vitro contraction model of electrically pulse-stimulated (EPS) C2C12 myotubes. By analyzing a subset of genes representing inflammatory, metabolic, and structural adaptation pathways, we could show that OA supplementation diminishes the EPS-provoked expression of inflammatory transcripts (interleukin 6, Il6; chemokine (C-X-C Motif) ligand 5, Cxcl5), and attenuated transcript levels of nuclear receptor subfamily 4 group A member 3 (Nr4A3), early growth response 1 (Egr1), activating transcription factor 3 (Atf3), and fast-oxidative MyHC-IIA isoform (Myh2). By contrast, OA had no suppressive effect on the pathogen-provoked inflammatory gene response in skeletal muscle cells, as demonstrated by stimulation of C2C12 myotubes with bacterial LPS. In addition, we observed a suppressive effect of OA on EPS-induced phosphorylation of AMP-activated protein kinase (AMPK), whereas EPS-triggered phosphorylation/activation of the mammalian target of rapamycin (mTOR) was not affected. Finally, we demonstrate that OA positively influences glycogen levels in EP-stimulated myotubes. Taken together, our results suggest that in skeletal muscle cells, OA modulates both the inflammatory and the metabolic reaction provoked by acute contraction. These results might have important clinical implications, specifically in cardiovascular and exercise medicine.

Acute exercise-induced irisin release in healthy adults: Associations with training status and exercise mode

Abstract There is a growing interest in exploring irisin response to acute exercise; however, the associations of acute exercise-induced irisin release with training status and exercise mode are not fully understood. This study was primarily designed to evaluate these associations. Sixteen healthy adults (8 trained versus 8 untrained) underwent a bout of cycling at 80% of maximal oxygen uptake (VO2max) for 50 min, with blood drawn pre-, 10-, and 180-min post-exercise. Another 17 healthy adults performed 2 bouts of graded exercise (cycling and running) until exhaustion on separate days using a randomized cross-over design, with blood taken pre-, 0-, 10-, and 60-min post-exercise. Circulating irisin, creatine kinase (CK), aspartate aminotransferase (AST), and myoglobin (Mb) were measured, and their respective areas under the curves (AUCs) were calculated. Irisin increased 10-min after 50 min of cycling at 80% of VO2max, while its changes from baseline to post-exercise and the amount of exercise-induced irisin release (presented as AUC) were comparable between trained and untrained adults (all P > .05). Irisin remained elevated 10-min post-exhausting running but decreased towards baseline 10-min post-exhausting cycling. Exhausting running induced an increase in irisin release for the whole course of exercise and recovery periods, but cycling did not. Acute exercise-induced irisin changes seemed not related to changes of CK, aspartate AST, and Mb in general. In conclusion, acute exercise-induced irisin release is not associated with training status but might be affected by training mode. Future studies are required to investigate which exercise mode might be most efficient in altering irisin.

Mid-term development of the right ventricle in competitive athletes:

Background Long-term intensive training induces physiological, morphological, and functional adaption of the athlete’s heart. Purpose To evaluate the development of athlete’s heart during a mid-term follow-up of competitive athletes using cardiac magnetic resonance (CMR). Material and Methods Eighteen competitive long-distance runners and triathletes (age 43 ± 13 years, 3 women) were prospectively examined in a longitudinal follow-up study 5.05 ± 0.6 years after baseline. CMR at 1.5-T was performed for functional and late gadolinium enhancement (LGE) imaging. Left ventricular (LV) and right ventricular (RV) end-diastolic volume (LVEDV, RVEDV) as well as ejection fraction (LVEF, RVEF), LV myocardial mass (LVMM), and atrial sizes were determined and compared to baseline in matched pairs statistics for paired difference. Results LVEDV (197 ± 38 mL vs. 196 ± 38 mL, paired difference −0.9 mL, P = 0.7) and LVEF (62 ± 7% vs. 62 ± 5%, paired difference 0.1%, P = 0.9) did not change during the follow-up period, whereas LVMM increased significantly (149 ± 31 g vs.164 ± 32 g, paired difference 14 g, P < 0.0001). RVEDV significantly increased from 221 ± 47 mL at baseline to 230 ± 52 mL (paired difference 10 mL, P = 0.0033). RVEF decreased from baseline 57 ± 8% to 53 ± 7% (paired difference −3%, P = 0.0234). Left atrial size showed no significant changes (24 ± 5 cm2 vs. 25 ± 6 cm2, paired difference 0.5 cm2, P = 0.17) and right atrial size increased significantly (30 ± 5 cm2 vs. 32 ± 4 cm2, paired difference 2 cm2, P = 0.0054). Conclusion This study supports the theory of ongoing remodeling in an athlete’s heart. Predominantly the right heart can further enlarge in a mid-term period. This response seems not linearly dependent on a steady, decreased, or increased training volume.

Correspondence (letter to the editor): Cycle Ergometry Is not a Reliable Parameter

We work in an institution that provides follow-up examinations for patients with coronary heart disease and wish to share the following observations. The authors rightly say that routine CT angiography is not indicated after coronary stent implantation. This is not only because the coronary stenoses diagnosed on CT are often not hemodynamically relevant. The more important reason is that monitoring stents is affected by their structure and size, among others. Further, in patients with known coronary heart disease the coronary vessels are often subject to severe calcification, which can substantially limit diagnostic accuracy. The consensus recommendations from the working groups of the German Cardiac Society/German Society of Pediatric Cardiology do not regard the indication as definite in asymptomatic and symptomatic patients after coronary intervention or bypass surgery, unless the question is only one of bypass graft patency (1). Regarding ergometry testing, our critical comment would be that the reported criterion of maximum exertion (85% of age-adjusted heart frequency) does not constitute a reliable parameter. The maximum individual heart frequency is subject to large variability and is furthermore affected by cardiac medication (2, 3). Furthermore, especially untrained elderly patients cannot achieve (cardiac) maximum exertion on the exercise bike because of deficits in the peripheral muscles. Exercise on a treadmill can be helpful in this setting as it typically reflects everyday activity more closely, in order to reach as high a degree of maximum exertion as possible and to improve the sensitivity of the exercise ECG, which is limited anyway. Furthermore, it is possible by using combinations with lactate testing and/or spiro-ergometry testing to establish more objective criteria for maximum exertion, but also to finesse the control of the recommended physical training.

WISSENSCHAFTLICHER BEITRAG: Liste der verbotenen Substanzen 2005 – Konsequenzen für die Behandlung von Leistungssportlern

Summary The modifications of the World Anti-Doping Agency (WADA) in the 2005 prohibited list – international standard causes some changes in the medical therapy of elite athletes. That applies not only to junior and high performance athletics, but also to senior competition athletics. Structurally, there is now a differentiation between substances and methods prohibited at all times (in- and out-of-competition) and substances and methods prohibited only in-competition. Moreover there is a differentiation between open lists and closed lists. There are three essential modifications to the content in contrast to the 2004 prohibited list. Beta-2-agonsits are now prohibited in- and out-of-competition. The use of these substances requires a therapeutic use exemption – standard application form (TUE). As an exception, Formoterol, Salbutamol, Salmeterol, and Terbutalin, when administered by inhalation require an abbreviated therapeutic use exemption (ATUE) only. Dermatological preparations of glucocorticosteroids in the form of ointments or creams are no longer prohibited. Administration of glucocorticosteroids by inhalation and non-systemic routes continues to require an ATUE. The therapy of chronic diseases, for example systemic therapy using insulin or cortisone, requires a TUE. Intravenous infusions are now listed as a prohibited method. Prohibition of intravenous infusions is not aimed at preventing their use for legitimate acute medical purposes.