Acute and long-term effects of blood flow restricted training on heat shock proteins and endogenous antioxidant systems.

Blood flow restricted exercise (BFRE) with low loads has been demonstrated to induce considerable stress to exercising muscles. Muscle cells have developed a series of defensive systems against exercise-induced stress. However, little is known about acute and long-term effects of BFRE training on these systems. Nine previously untrained females trained low-load BFRE and heavy load strength training (HLS) on separate legs and on separate days to investigate acute and long-term effects on heat shock proteins (HSP) and endogenous antioxidant systems in skeletal muscles. BFRE and HLS increased muscle strength similarly by 12 ± 7% and 12 ± 6%, respectively, after 12 weeks of training. Acutely after the first BFRE and HLS exercise session, αB-crystallin and HSP27 content increased in cytoskeletal structures, accompanied by increased expression of several HSP genes. After 12 weeks of training, this acute HSP response was absent. Basal levels of αB-crystallin, HSP27, HSP70, mnSOD, or GPx1 remained unchanged after 12 weeks of training, but HSP27 levels increased in the cytoskeleton. Marked translocation of HSP to cytoskeletal structures at the commencement of training indicates that these structures are highly stressed from BFRE and HLS. However, as the muscle gets used to this type of exercise, this response is abolished.

Strength training improves cycling performance, fractional utilization of VO2max and cycling economy in female cyclists.

The purpose of this study was to investigate the effect of adding heavy strength training to well-trained female cyclists' normal endurance training on cycling performance. Nineteen female cyclists were randomly assigned to 11 weeks of either normal endurance training combined with heavy strength training (E+S, n = 11) or to normal endurance training only (E, n = 8). E+S increased one repetition maximum in one-legged leg press and quadriceps muscle cross-sectional area (CSA) more than E (P < 0.05), and improved mean power output in a 40-min all-out trial, fractional utilization of VO2 max and cycling economy (P < 0.05). The proportion of type IIAX-IIX muscle fibers in m. vastus lateralis was reduced in E+S with a concomitant increase in type IIA fibers (P < 0.05). No changes occurred in E. The individual changes in performance during the 40-min all-out trial was correlated with both change in IIAX-IIX fiber proportion (r = -0.63) and change in muscle CSA (r = 0.73). In conclusion, adding heavy strength training improved cycling performance, increased fractional utilization of VO2 max , and improved cycling economy. The main mechanisms behind these improvements seemed to be increased quadriceps muscle CSA and fiber type shifts from type IIAX-IIX toward type IIA.

Strength training improves performance and pedaling characteristics in elite cyclists.

The purpose was to investigate the effect of 25 weeks heavy strength training in young elite cyclists. Nine cyclists performed endurance training and heavy strength training (ES) while seven cyclists performed endurance training only (E). ES, but not E, resulted in increases in isometric half squat performance, lean lower body mass, peak power output during Wingate test, peak aerobic power output (W(max)), power output at 4 mmol L(-1)[la(-)], mean power output during 40-min all-out trial, and earlier occurrence of peak torque during the pedal stroke (P < 0.05). ES achieved superior improvements in W(max) and mean power output during 40-min all-out trial compared with E (P < 0.05). The improvement in 40-min all-out performance was associated with the change toward achieving peak torque earlier in the pedal stroke (r = 0.66, P < 0.01). Neither of the groups displayed alterations in VO2max or cycling economy. In conclusion, heavy strength training leads to improved cycling performance in elite cyclists as evidenced by a superior effect size of ES training vs E training on relative improvements in power output at 4 mmol L(-1)[la(-)], peak power output during 30-s Wingate test, W(max), and mean power output during 40-min all-out trial.

Reliable determination of training-induced alterations in muscle fiber composition in human skeletal muscle using quantitative polymerase chain reaction.

Determination of muscle fiber composition in human skeletal muscle biopsies is often performed using immunohistochemistry, a method that tends to be both time consuming, technically challenging, and complicated by limited availability of tissue. Here, we introduce quantitative reverse transcriptase polymerase chain reaction (qRT-PCR)-based Gene-family profiling (GeneFam) of myosin heavy chain (MyHC) mRNA expression as a high-throughput, sensitive, and reliable alternative. We show that GeneFam and immunohistochemistry result in similar disclosures of alterations in muscle fiber composition in biopsies from musculus vastus lateralis and musculus biceps brachii of previously untrained young women after 12 weeks of progressive strength training. The adaptations were evident as (a) consistent increases in MyHC2A abundance; (b) consistent decreases in MyHC2X abundance; and (c) consistently stable MyHC1 abundance, and were not found using traditional reference gene-based qRT-PCR analyses. Furthermore, muscle fiber composition found using each of the two approaches was correlated with each other (r = 0.50, 0.74, and 0.78 for MyHC1, A, and X, respectively), suggesting that GeneFam may be suitable for ranking of individual muscle phenotype, particularly for MyHC2 fibers. In summary, GeneFam of MyHC mRNA resulted in reliable assessment of alterations in muscle fiber composition in skeletal muscle of previously untrained women after 12 weeks of strength training.

Irisin and FNDC5: effects of 12-week strength training, and relations to muscle phenotype and body mass composition in untrained women.

PURPOSE: To investigate the effects of strength training on abundances of irisin-related biomarkers in skeletal muscle and blood of untrained young women, and their associations with body mass composition, muscle phenotype and levels of thyroid hormones. METHODS: Eighteen untrained women performed 12 weeks of progressive whole-body heavy strength training, with measurement of strength, body composition, expression of irisin-related genes (FNDC5 and PGC1α) in two different skeletal muscles, and levels of serum-irisin and -thyroid hormones, before and after the training intervention. RESULTS: The strength training intervention did not result in changes in serum-irisin or muscle FNDC5 expression, despite considerable effects on strength, lean body mass (LBM) and skeletal muscle phenotype. Our data indicate that training affects irisin biology in a LBM-dependent manner. However, no association was found between steady-state serum-irisin or training-associated changes in serum-irisin and alterations in body composition. FNDC5 expression was higher in m.Biceps brachii than in m.Vastus lateralis, with individual expression levels being closely correlated, suggesting a systemic mode of transcriptional regulation. In pre-biopsies, FNDC5 expression was correlated with proportions of aerobic muscle fibers, a relationship that disappeared in post-biopsies. No association was found between serum-thyroid hormones and FNDC5 expression or serum-irisin. CONCLUSION: No evidence was found for an effect of strength training on irisin biology in untrained women, though indications were found for a complex interrelationship between irisin, body mass composition and muscle phenotype. FNDC5 expression was closely associated with muscle fiber composition in untrained muscle.

Effects of 12 weeks of block periodization on performance and performance indices in well-trained cyclists.

The purpose of this study was to compare the effects of two different methods of organizing endurance training in trained cyclists during a 12-week preparation period. One group of cyclists performed block periodization (BP; n = 8), wherein every fourth week constituted five sessions of high-intensity aerobic training (HIT), followed by 3 weeks of one HIT session. Another group performed a more traditional organization (TRAD; n = 7), with 12 weeks of two weekly HIT sessions. The HIT was interspersed with low-intensity training (LIT) so that similar total volumes of both HIT and LIT were performed in the two groups. BP achieved a larger relative improvement in VO2max than TRAD (8.8 ± 5.9% vs 3.7 ± 2.9%, respectively, < 0.05) and a tendency toward larger increase in power output at 2 mmol/L [la(-)] (22 ± 14% vs 10 ± 7%, respectively, P = 0.054). Mean effect size (ES) of the relative improvement in VO2max , power output at 2 mmol/L [la(-)], hemoglobin mass, and mean power output during 40-min all-out trial revealed moderate superior effects of BP compared with TRAD training (ES range was 0.62-1.12). The present study suggests that BP of endurance training has superior effects on several endurance and performance indices compared with TRAD.

Block periodization of high-intensity aerobic intervals provides superior training effects in trained cyclists.

The purpose of this study was to compare the effect of two different methods of organizing endurance training in trained cyclists. One group of cyclists performed block periodization, wherein the first week constituted five sessions of high-intensity aerobic training (HIT), followed by 3 weeks of one weekly HIT session and focus on low-intensity training (LIT) (BP; n = 10, VO2max = 62 ± 2 mL/kg/min). Another group of cyclists performed a more traditional organization, with 4 weeks of two weekly HIT sessions interspersed with LIT (TRAD; n = 9, VO2max = 63 ± 2 mL/kg/min). Similar volumes of both HIT and LIT was performed in the two groups. While BP increased VO2max , peak power output (Wmax) and power output at 2 mmol/L [la(-)] by 4.6 ± 3.7%, 2.1 ± 2.8%, and 10 ± 12%, respectively (P < 0.05), no changes occurred in TRAD. BP showed relative improvements in VO2max compared with TRAD (P < 0.05). Mean effect size (ES) of the relative improvement in VO2max , Wmax , and power output at 2 mmol/L [la(-)] revealed large to moderate effects of BP training compared with TRAD training (ES = 1.34, ES = 0.85, and ES = 0.71, respectively). The present study suggests that block periodization of training provides superior adaptations to traditional organization during a 4-week endurance training period, despite similar training volume and intensity.

Per-unit-living tissue normalization of real-time RT-PCR data in ischemic rat hearts.

In studies of gene expression in acute ischemic heart tissue, internal reference genes need to show stable expression per-unit-living tissue to hinder dead cells from biasing real-time RT-PCR data. Until now, this important issue has not been appropriately investigated. We hypothesized that the expression of seven internal reference genes would show stable per-unit-living tissue expression in Langendorff-perfused rat hearts subjected to ischemia-reperfusion. This was found for cyclophilin A, GAPDH, RPL-32, and PolR2A mRNA, with GAPDH showing the highest degree of stability (R = 0.11), suggesting unchanged rates of mRNA transcription in live cells and complete degradation of mRNA from dead cells. The infarct size-dependent degradation of GAPDH was further supported by a close correlation between changes in GAPDH mRNA and changes in RNA quality measured as RNA integrity number (R = 0.90, P < 0.05). In contrast, ß-actin and 18S rRNA showed stable expression per-unit-weight tissue and a positive correlation with infarct size (R = 0.61 and R = 0.77, P < 0.05 for both analyses). The amount of total RNA extracted per-unit-weight tissue did not differ between groups despite wide variation in infarct size (7.1-50.1%). When ß-actin expression was assessed using four different normalization strategies, GAPDH and geNorm provided appropriate per-unit-living expression, while 18S and total RNA resulted in marked underestimations. In studies of ischemic tissues, we recommend using geometric averaging of carefully selected reference genes for normalization of real-time RT-PCR data. A marked shift in the mRNA/rRNA ratio renders rRNA as useless for normalization purposes.

Hypotonic stress activates BK channels in clonal kidney cells via purinergic receptors, presumably of the P2Y subtype.

AIM: Membrane stretch due to cell swelling may cause a minute leakage of adenosine triphosphate (ATP) that stimulates endogenous purinergic receptors. The following elevation of the cytosolic-free Ca(2+) concentration ([Ca(2+)](i)) may then participate in cell volume regulation. The aim of the present study was to test if purinergic receptors and large conductance Ca(2+) activated K(+) (BK) channels are activated in response to hypotonic stress in clonal kidney cells (Vero cells). METHODS: The methods used are fura-2 microfluorometry, cell-attached patch clamp and reverse-transcriptase polymerase chain reaction (RT-PCR). METHODS: Subjecting cells to hypotonic stress for 10 s by exposure to a solution with 45% reduced osmolality induced a transient rise in [Ca(2+)](i). This response persisted in virtually Ca(2+)-free extracellular solution, demonstrating that Ca(2+) was mainly released from intracellular stores. The hypotonically induced elevation of [Ca(2+)](i) was completely inhibited by the P2 receptor antagonists suramine (100 microM) and pyridoxalphosphate-6-azophenyl-2'4'-disulphonate (PPADS; 20 microM), indicating that extracellular ATP is crucial for the [Ca(2+)](i) increase. RT-PCR revealed the expression of mRNA for P2Y(1) receptors in Vero cells. The putatively selective P2Y(1) antagonist PPADS did completely block Ca(2+) responses to both ATP and hypotonic stress, suggesting that P2Y(1) receptors are mediating the response. Furthermore, patch clamp recordings in cell-attached configuration revealed that BK channels are activated in response to hypotonic stress. conclusion: Vero cells express functional purinergic receptors, presumably of the P2Y(1) subtype. These receptors are responsible for the elevation of [Ca(2+)](i) evoked by hypotonic stress. The concurrent activation of BK channels permits K(+) efflux that may contribute to regulatory volume decrease.