Martin Schönfelder

High-intensity interval training is not superior to other forms of endurance training during cardiac rehabilitation:

Background High-intensity interval training has recently emerged as superior to continuous endurance training in cardiac rehabilitation upon other training regimes. Individually tailored continuous endurance training and pyramid training could induce comparable effects on peak work capacity as high intensity interval training. Design A prospective, randomized study. Methods Effects of the following isocaloric cycle ergometer protocols on peak work capacity have been assessed in patients with coronary artery disease (n = 60) during 6 weeks of outpatient cardiac rehabilitation, i.e. 18 supervised sessions of exercise training: (1) continuous endurance training (n = 20): 33 min at 65–85% peak heart rate; (2) high intensity interval training (n = 20): 4 × 4 min intervals at 85–95% peak heart rate, each followed by 3 min of active recovery at 60–70% peak heart rate; (3) pyramid training (n = 20): 3 × 8 min of stepwise load increase and subsequent decrease from 65–95–65% peak heart rate, supplemented by 2 min recovery at 60–70% peak heart rate between pyramids. All protocols were preceded by 5 min of warm-up and followed by 5 min cool-down at 60–70% peak heart rate. Results Attendance during exercise sessions was 99.2%. There were significant increases in peak work capacity of comparable magnitude in all three training groups (begin vs. end: continuous endurance training: 136.0 ± 49.6 W vs. 163.4 ± 60.8 W (21.1 ± 8.5%); high-intensity interval training: 141.0 ± 60.4 W vs. 171.1 ± 69.8 W (22.8 ± 6.6%); pyramid training: 128.7 ± 50.6 W vs. 158.5 ± 57.9 W (24.8 ± 10.8%); within groups all p < 0.001; between groups, p = not significant). Conclusion Endurance training protocols assessed in this study all led to significant increases in peak work capacity of comparable magnitude. Our findings suggest that these protocols can be used interchangeably, which will lead to further individualization of exercise prescription and may therefore result in improved adherence to lifelong behavioural changes.

Androgen-dependent morphology of prostates and seminal vesicles in the Hershberger Assay: Evaluation of immunohistochemical and morphometric parameters*

The aim of this study was to evaluate androgen-like effects using immunohistochemical and morphometric methods. Therefore, orchiectomized Wistar rats (n > or = 13) were treated s.c. with 1 mg/kg bw/day testosterone propionate (TP) for 7 days and compared to orchiectomized rats without TP substitution (OX) and to an untreated intact control group. Sections obtained from prostates and seminal vesicles were stained with polyclonal and monoclonal antibodies against the androgen receptor (AR) and assessed densitometrically (intensity of the immunoreaction) and morphometrically (epithelial height, luminal area). TP caused an enhancement of staining intensity and an increase in organ weights, epithelial height and luminal area. The use of proliferation markers (PCNA, MIB-5) showed also a highly significant increase of immunoreactive cells in TP-substituted orchiectomized rats compared with the OX group. Based on the present data, the densitometric analysis of AR-immunoreactivity as well as the assessment of proliferation markers, epithelial height and luminal area proved to be sensitive parameters for the evaluation of androgen effects on prostates and seminal vesicles. In further studies these parameters will be used to test several industrial xenooestrogens as well as phytooestrogens on their possible androgenic capacity.

Scientific Comparison of Different Online Heart Rate Monitoring Systems

Recent technical development focused on real-time heart rate monitoring instead of postexercise evaluation of recorded data. There are several systems on the market that allow direct and real-time monitoring of several individuals at the same time. The present study compared the systems of Polar, Acentas, Activio, and Suunto in a field test with twelve subjects regarding failure quota, operating distance, and ECG validity. Moreover, the installation and use of software and hardware were evaluated with a quality rating system. Chest belts were evaluated with a questionnaire, too. Overall the system of Acentas reached the best mark of all systems, but detailed results showed that every system has its advantages and disadvantages depending on using purpose, location, and weather. So this evaluation cannot recommend a single system but rather shows strength and weakness of all systems and additionally can be used for further system improvements.

Gene expression profiling in human whole blood samples after controlled testosterone application and exercise

Doping with anabolic agents is regulated within a number of sports. Testosterone and its functional analogs are popular compounds for increasing muscle mass, physical performance, recovery, and reducing body fat. While routine tests for anabolic drugs exist (e.g. hair, urine, and blood analysis), the aim of the present study is to determine specific gene expression profiles (induced by testosterone and exercise) which may be used as effective biomarkers to determine the use of anabolic drugs. In this study, whole blood samples of 19 male volunteers were analyzed by semi-quantitative real-time polymerase chain reaction (RT-PCR) for gene expression profiles in the context of exercise and transdermal testosterone application (1.5 mg/kg body weight). The hormone application was monitored by urine and saliva analysis for testosterone. Both urinary and saliva levels indicate that transdermal testosterone application leads to an increase of testosterone, especially after exercise. RT-PCR results showed a clear variation in the expression of target genes as well as established housekeeping genes. Only one of the nine common housekeeping genes, cyclophilin b (PPIB), appears to be independent of both exercise and testosterone. Out of 14 candidate genes, five are unregulated; all others were more or less influenced by the mentioned variables. Only interleukin-6 appeared to be exclusively dependent on long-term testosterone application. This study indicates that many genes are not influenced by testosterone alone while exercise modulates gene expression in whole blood samples. As such, exercise must be considered when validating gene expression techniques for doping analysis.

The dependence of autologous chondrocyte transplantation on varying cellular passage, yield and culture duration

Matrix-assisted chondrocyte transplantation (m-ACI) still lacks any standardization in its execution in terms of cell passage (P), cell yield (C) and in vitro membrane-holding time (T). It was the goal of this study to analyze the effect of shifting cell culture parameters (P, C, T) on the in vitro as well as in vivo effort of a regulated animal m-ACI. Autologous rabbit knee articular chondrocytes were seeded within bilayer collagen I/III 3-D matrices in variation of P, C and T. Each time, 2 PCT-identical by 2 PCT-identical cell-matrix-constructs (CMC)/animal were created. Simultaneously 2 (PCT-distinct) were re-implanted (CMC-e) autologous into artificial trochlear pristine chondral defects in vivo to remain for 12 weeks while the remaining 2 were harvested (CMC-i) for immediate in vitro analysis at the time of transplantation of their identical twins. mRNA of both, CMC-e regenerates and CMC-i membranes, was analyzed for Collagen-1,-2,-10, COMP, Aggrecan, Sox9 expression by use of a mixed linear model, multiple regression analysis. Generally, CMC-i values were higher than CMC-e values for differentiation targets; the opposite was true for dedifferentiation targets. Regarding individual gene expression, in vivo regenerate cell-matrix properties were significantly dependent on initial cell-matrix in vitro values as a sign of linearity. The parameter membrane-holding time (T) had strongest effects on the resulting mRNA expression with slightly less impact of the parameter passage (P), whereas cell yield (C) had clearly less effects. Noting differences between in vitro and in vivo data, in general, optimal expression patterns concerning chondrogenic differentiation were achieved by few passages, medium cellular yield, short membrane-holding time. Clinical m-ACI may benefit from optimal orchestration of the cell culture parameters passage, yield and time.

Gene Expression in Hair Follicle Dermal Papilla Cells after Treatment with Stanozolol

Doping with anabolic agents is a topic in sports where strength is crucial, e.g. sprinting, weight lifting and many more. Testosterone and its functional analogs are the drugs of choice taken as pills, creams, tape or injections to increase muscle mass and body performance, and to reduce body fat. Stanozolol (17β-hydroxy-17α-methyl-5α-androst-2-eno[3,2c]pyrazol) is a testosterone analogue with the same anabolic effect like testosterone but its ring structure makes it possible to take it orally. Therefore, stanozolol is one of the most frequently used anabolic steroids. Common verification methods for anabolic drugs exist, identifying the chemicals in tissues, like hair or blood samples. The idea of this feasibility study was to search for specific gene expression regulations induced by stanozolol to identify the possible influence of the synthetically hormone on different metabolic pathways. Finding biomarkers for anabolic drugs could be supportive of the existing methods and an additional proof for illegal drug abuse. In two separate cell cultures, human HFDPC (hair follicle dermal papilla cells) from a female and a male donor were treated with stanozolol. In the female cell culture treatment concentrations of 0 nM (control), 1 nM, 10 nM and 100 nM were chosen. Cells were taken 0 h, 6 h, 24 h and 48 h after stimulation and totalRNA was extracted. Learning from the results of the pilot experiment, the male cell culture was treated in 10 nM and 100 nM concentrations and taken after 0 h, 6 h, 24 h and 72 h. Using quantitative real-time RT-PCR expression of characteristics of different target genes were analysed. Totally 13 genes were selected according to their functionality by screening the actual literature and composed to functional groups: factors of apoptosis regulation were Fas Ligand (FasL), its receptor (FasR), Caspase 8 and Bcl-2. Androgen receptor (AR) and both estrogen receptors (ERα, ERβ) were summarized in the steroid receptor group. The growth factor group included the insulin like growth factor receptor (IGF1R) and growth hormone receptor (GHR). Fibroblast growth factor 2 (FGF2) and keratinocyte growth factor (FGF7) were summarized in the hair cycle factor group. 5α-Steroidreductases (SRD5A1, SRD5A2) represented the enzyme group. Three reference genes were taken for relative quantification: ubiquitin (UBQ), glycerinaldehyde-3-phsophate-dehydrogenase (GAPDH), and β-actin (ACTB). In cell culture 1 AR, FasR, FGF2 showed significant regulations within one treatment time, significant gene expressions over time were analysed for Caspase 8. In cell culture 2 AR, FasR and SRD5A2 were significantly regulated within one treatment time. In this feasibility study first biomarker for a screening pattern of anabolic agents could be identified providing the rationality to investigate modified, metabolic pathways in the whole hair follicle.

The xenoestrogen bisphenol A in the Hershberger assay: androgen receptor regulation and morphometrical reactions indicate no major effects.

We evaluated androgen-like effects of bisphenol A (BPA) using orchiectomized Wistar rats. Animals were treated p.o. either with vehicle or with 3, 50, 200, 500 mg/kgbw/day BPA (n=13) for 7 days. One group was treated s.c. with 1mg/kgbw/day testosterone propionate (TP). Flutamide (FL) (3mg/kgbw/day, p.o.) was used to antagonize androgen effects of the suprapharmacological dose (500 mg/kgbw/day) of BPA. Androgen-like effects of BPA on prostates and seminal vesicles were assessed by the Hershberger assay, densitometric analysis of androgen receptor (AR) immunoreactivity, cell proliferation-index and a morphometric analysis. Absolute weights of prostates and seminal vesicles were not increased by BPA, whereas the relative weights were increased at higher doses of BPA, most likely due to a decrease in body weight. Staining intensity for AR immunoreactivity was increased at low but not at higher doses of BPA in comparison to the orchiectomized rats. BPA at all doses tested did not cause an increase of the cell proliferation-index. Epithelial height and glandular luminal area were increased by low doses of BPA, whereas higher doses caused a decrease of these parameters. The data provide evidence that BPA does not exert major androgenic effects.

Potential detection of low-dose transdermal testosterone administration in blood, urine, and saliva.

Administration of low amounts of endogenous hormones - so-called micro-dosages - are supposed to represent a major challenge in doping analysis. To model such a situation, we have studied transdermal administrations of 2.4 mg/24 h testosterone patches and examined various steroid concentrations in blood, urine, and saliva of 11 volunteers. Multiple samples were collected at t = 0, 3, 6, 9, 24, 48, and 72 h in four different phases, i.e., all combinations with/without physical exercise and with/without testosterone. Testosterone was analyzed by enzyme-linked-immuno-assay as well as by mass spectrometry and validated in an accredited anti-doping laboratory. Circadian controls with and without exercise did not provoke prominent alterations of whole, free, and salivary testosterone. Testosterone application for 24 h led to a significant (all p < 0.001) mean increase above controls: total testosterone (median: 5.2 vs. 8.0 ng/mL), free testosterone (median: 11.3 vs. 15.6 pg/mL), and salivary testosterone (median: 62.4 vs. 99.9 pg/mL). Additionally, all three testosterone measurements indicated significant correlations to each other (all r > 0.538, all p < .001). Circadian-matching showed peaking testosterone values after 6 h and 9 h, reaching highest augmentation up to 252.6 ± 123.5% in saliva after 9 h. After removal of the testosterone patch, all testosterone levels in blood, saliva, and urine returned to baseline within 24 h. Different techniques of hormone detection (enzyme-linked immunosorbent assay (ELISA), gas chromatography-tandem mass spectrometry (GC-MS/MS), liquid chromatography-tandem mass spectrometry (LC-MS/MS)) indicated significant correlations. Results indicate that saliva, blood, and urine exhibit comparable hormone augmentation during micro-dose testosterone application, indicating a possible consideration in future doping analysis. The inter-individual variability was high in all biofluids, requiring the use of an individual biological passport rather than statistical values. Copyright

Does exercise training impact clock genes in patients with coronary artery disease and type 2 diabetes mellitus

Background Recent findings revealed negative effects of deregulated molecular circadian rhythm in coronary artery disease (CAD) and type 2 diabetes mellitus (T2DM). Physical exercise training (ET) has been shown to promote anti-diabetic and anti-atherogenic responses in skeletal muscle of these patients, but the role of the circadian clock-machinery remains unknown. This study investigated whether mRNA expression of clock genes in skeletal muscle of CAD and T2DM patients is influenced by physical ET intervention. Methods Nineteen patients with CAD and T2DM (age 64 ± 5 years) were randomised to either six months of ET (four weeks of in-hospital ET followed by a five-month ambulatory programme) or usual care. At the beginning of the study, after four weeks and after six months parameters of metabolic and cardiovascular risk factors, and physical exercise capacity were assessed. Gene expression was measured in skeletal muscle biopsies by quantitative real-time polymerase chain reaction (PCR). Results A selection of clock genes and associated components (circadian locomoter output cycle kaput protein (CLOCK), period (PER) 1, cryptochrome (CRY) 2 and aminolevulinate-deltA-synthase-1 (ALAS1)) was reliably measured and used for further analysis. A time-dependent effect in gene expression was observed in CLOCK (p = 0.013) and a significant interaction between time and intervention was observed for ALAS1 (p = 0.032; p = 0.014) as a result of ET. Conclusion This is the first study to analyse clock gene expression in skeletal muscles of patients with CAD and T2DM participating in a long-lasting exercise intervention. ET, as one of the cornerstones in prevention and rehabilitation of CAD and T2DM, exerts no effects on CLOCK genes but meaningful effects on the clock-associated gene ALAS1.

Alpine Skiing as Winter-time High-intensity Training

Introduction To counteract the winter activity deficit, we set out to analyze cardiorespiratory and metabolic responses of two high-intensity training (HIT) protocols during alpine skiing (AS), cross-country skiing (XCS), and indoor cycling (IC) and the effects of sex, age, and fitness level in this comparison. Methods Nineteen healthy subjects (two age and fitness groups, both sexes) performed AS, XCS, and IC with measurements of oxygen uptake (V˙O2), energy expenditure (EE), HR, lactate, blood glucose and rate of perceived exertion, determined during 4 min of continuous HIT (HITc: 90% HRmax for XCS and IC or short turn skiing during AS) or 10-min intermittent HIT [HITint: 5 × 1 min high intensity (>90% HRmax or short turn skiing), 1 min active recovery]. Results During all three exercise modes and irrespective of HIT protocols, sex, age, and fitness, participants were able to reach exercise intensities >90% HRmax and >84% V˙O2max. In all exercise modes 10-min of HITint with a 10-min postexercise O2 consumption phase resulted in greater mean EE per minute compared to 4-min HITc with 10 min postexercise O2 consumption. When applying the same HIT loading and recovery pattern to all three exercise modes, EE during approximately 1:15 h of AS was equivalent to about 1:00 h of either XCS or IC. Conclusions Across all exercise modes and HIT protocols, high cardiorespiratory and metabolic responses were achieved regardless of age, sex, or fitness. EE during AS can be maximized by choosing the skiing mode “short turn skiing” in combination with an HITint to prolong the duration of continuous high-intensity loading during each descent. Therefore, all exercise modes and both HIT protocols are applicable and feasible in a broad spectrum of healthy subjects.