Reduction in plasma leucine after sprint exercise is greater in males than in females.

There is a pronounced gender difference in the accumulation of plasma ammonia after sprint exercise. Ammonia is a key intermediate in amino acid metabolism, which implies that gender-related differences in plasma and muscle amino acid concentrations after sprint exercise exist. To study this, three bouts of 30-s sprint exercise were performed by healthy females (n=8) and males (n=6). Blood leucine and muscle leucine were collected over the exercise period. Basal arterial plasma and skeletal muscle leucine were 40% higher in males than females (P<0.010 and P<0.020). Plasma, but not muscle, leucine decreased by sprint exercise and more so in males than females (g × t: P=0.025). Increase in ammonia was higher in males than females in both plasma and muscle (g × t: P<0.001 and P=0.003). An opposite pattern was shown for plasma glutamine, where an increase was found in females (P<0.001), but not in males. In conclusion, the lower plasma ammonia after sprint exercise in females seems to be explained by a lower accumulation of ammonia in skeletal muscle and by a buffering of ammonia in the form of glutamine in females. The greater reduction in plasma leucine in males seems to be related to their greater increase in muscle ammonia after sprint exercise.

Expression of both oestrogen receptor alpha and beta in human skeletal muscle tissue.

There are two oestrogen receptors (ERs), ERalpha and ERbeta. ERbeta protein is expressed in human skeletal muscle in the nuclei of both myofibres and endothelial cells, whether ERalpha protein is present in this tissue is unknown. We studied the expression of ERalpha protein in human skeletal muscle biopsies taken from vastus lateralis from four men, four women, two children and two postmenopausal women. Immunohistochemistry was used to determine the proportions of nuclei that were positively stained for ERalpha, the proportion of ERalpha-positive nuclei located in the muscle fibres and in capillaries and to test for possible co-expression of ERalpha and ERbeta. Both ERs were expressed in all subjects. Of all nuclei, 63% stained for ERalpha with no sex difference. ERalpha was localised both in myofibres and in endothelial cells of the capillaries, 25% of the ERalpha-positive nuclei were located in the capillaries. ERalpha and ERbeta were generally expressed in the same nuclei. The present study shows for the first time the expression of ERalpha protein in human skeletal muscle independently of age and sex. These results might improve understanding of the physiological role of oestrogen in human skeletal muscle and raise new questions about activation of ERs in skeletal muscle.

Muscle oxidative capacity and work performance after training under local leg ischemia.

Healthy young men executed supine one-legged cycle training four times per week for 4 wk with legs and the cycle ergometer inside a pressure chamber, the opening of which was sealed by a rubber membrane at the level of the crotch. Each training session started by training one leg under ischemic conditions induced by increased chamber pressure (50 mmHg) at the highest intensity tolerable for 45 min. Then the other leg was trained with the same power profile but normal atmospheric chamber pressure. Before and after the training period, both legs executed one-legged exercise tests under both normal and increased chamber pressure and muscle biopsies were taken from the vastus lateralis. Ischemic training increased performance more than normal training, the difference being greater for exercise executed under ischemic conditions. The difference in performance increase between the legs was paralleled by a greater muscle citrate synthase activity in the ischemically than in the normally trained leg.

Sprint training, in vitro and in vivo muscle function, and myosin heavy chain expression.

Sprint training represents the condition in which increases in muscle shortening speed, as well as in strength, might play a significant role in improving power generation. This study therefore aimed to determine the effects of sprint training on 1) the coupling between myosin heavy chain (MHC) isoform expression and function in single fibers, 2) the distribution of MHC isoforms across a whole muscle, and 3) in vivo muscle function. Seven young male subjects completed 6 wk of training (3-s sprints) on a cycle ergometer. Training was without effect on maximum shortening velocity in single fibers or in the relative distribution of MHC isoforms in either the soleus or the vastus lateralis muscles. Electrically evoked and voluntary isometric torque generation increased (P < 0.05) after training in both the plantar flexors (+8% at 50 Hz and +16% maximal voluntary contraction) and knee extensors (+8% at 50 Hz and +7% maximal voluntary contraction). With the shortening potential of the muscles apparently unchanged, the increased strength of the major lower limb muscles is likely to have contributed to the 7% increase (P < 0.05) in peak pedal frequency during cycling.

Sprint training effects on muscle myoglobin, enzymes, fiber types, and blood lactate.

The purpose of this study was to determine if changes in intra-muscular myoglobin concentration accompany histochemical and enzymatic adaptations to supra-maximal exercise training. Subjects were assigned to either a training group (N = 11), who trained 2 to 3 times weekly for 6 wk, or a control group (N = 6). Training progressed from two 15-s and two 30-s "all-out" sprints on a cycle ergometer during week 1 to six 15-s and six 30-s bouts per session during week 6. The Wingate test was performed before and after the 6 wk, but performance variables were not changed in either group. In the training group, peak lactate after the Wingate test was significantly higher after training. No significant changes in enzyme activities, myoglobin concentration, or fiber-type frequency were observed in the control group. In contrast, in the training group, the percent fast twitch oxidative fibers increased, myoglobin decreased, and both citrate synthase and phosphofructokinase activities increased (P less than 0.05). The results suggest that muscle myoglobin concentration is not increased by 6 wk of supra-maximal exercise training and that such training induces cellular adaptations without accompanying performance changes. Alternatively, the Wingate test is not a sensitive test of adaptations to the training.

Sprint exercise enhances skeletal muscle p70S6k phosphorylation and more so in women than in men.

AIM: Sprint exercise is characterized by repeated sessions of brief intermittent exercise at a high relative workload. However, little is known about the effect on mTOR pathway, an important link in the regulation of muscle protein synthesis. An earlier training study showed a greater increase in muscle fibre cross-sectional area in women than men. Therefore, we tested the hypothesis that the activation of mTOR signalling is more pronounced in women than in men. Healthy men (n=9) and women (n=8) performed three bouts of 30-s sprint exercise with 20-min rest in between. METHODS: Multiple blood samples were collected over time, and muscle biopsy specimens were obtained at rest and 140 min after the last sprint. RESULTS: Serum insulin increased by sprint exercise and more so in women than in men [gender (g) × time (t)]: P=0.04. In skeletal muscle, phosphorylation of Akt increased by 50% (t, P=0.001) and mTOR by 120% (t, P=0.002) independent of gender. The elevation in p70S6k phosphorylation was larger in women (g × t, P=0.03) and averaged 230% (P=0.006) as compared to 60% in men (P=0.04). Phosphorylation rpS6 increased by 660% over time independent of gender (t, P=0.003). Increase in the phosphorylation of p70S6k was directly related to increase in serum insulin (r=0.68, P=0.004). CONCLUSION: It is concluded that repeated 30-s all-out bouts of sprint exercise separated by 20 min of rest increases Akt/mTOR signalling in skeletal muscle. Secondly, signalling downstream of mTOR was stronger in women than in men after sprint exercise indicated by the increased phosphorylation of p70S6k.

Higher proportion of fast-twitch (type II) muscle fibres in idiopathic inflammatory myopathies - evident in chronic but not in untreated newly diagnosed patients.

OBJECTIVE: Polymyositis and dermatomyositis are idiopathic, inflammatory myopathies characterized by proximal muscle fatigue. Conventional immunosuppressive treatment gives a variable response. Biopsies from chronic patients display a low proportion type I and a high proportion of type II muscle fibres. This raised a suspicion that the low proportion of type I fibres might play a role in the muscle fatigue. AIM: To investigate whether the muscle fibre attributes evident in chronic myositis are characteristic for the polymyositis and dermatomyosistis diseases themselves. METHODS: Muscle biopsies were obtained from thigh muscle from untreated patients (n = 18), treated responders (n = 14) and non-responders (n = 6) and from healthy controls (n = 11), respectively. For clinical evaluations, creatine kinase, functional index of myositis and cumulative dose of cortisone were established. RESULTS: Chronic patients had a lower proportion of type I fibres and a higher proportion of type II fibres compared to untreated myositis patients and healthy controls. Fibre cross-sectional area (CSA) did not differ between patients and healthy individuals but all women had a 20% smaller type II fibre CSA compared to men. CONCLUSIONS: Untreated polymyositis and dermatomyositis patients and healthy controls have a different fibre type composition than chronic polymyositis and dermatomyositis patients. Fibre CSA did not differ between healthy controls or any of the patient groups. A low proportion of oxidative muscle fibres can therefore be excluded as a contributing factor causing muscle fatigue at disease onset and the gender difference should be taken into consideration when evaluating fibre CSA in myositis.

Greater growth hormone and insulin response in women than in men during repeated bouts of sprint exercise.

AIM: In a previous study, sprint training has been shown to increase muscle cross-sectional area in women but not in men [Eur J Appl Physiol Occup Physiol 74 (1996) 375]. We hypothesized that sprint exercise induces a different hormonal response in women than in men. Such a difference may contribute to explaining the observed gender difference in training response. METHOD: Metabolic and hormonal response to three 30-s sprints with 20-min rest between the sprints was studied in 18 physically active men and women. RESULTS: Accumulation of blood lactate [interaction term gender (g) x time (t): P = 0.022], and plasma ammonia (g x t: P < 0.001) after sprint exercise was greater in men. Serum insulin increased after sprint exercise more so in women than in men (g x t: P = 0.020), while plasma glucose increased in men, but not in women (g x t: P < 0.001). Serum growth hormone (GH) increased in both women and men reaching similar peak levels, but with different time courses. In women the peak serum GH level was observed after sprint 1, whereas in men the peak was observed after sprint 3 (g x t; P < 0.001). Serum testosterone tended to decrease in men and increase in women (g x t: P = 0.065). Serum cortisol increased approx. 10-15% after sprint exercise, independent of gender (time: P = 0.005). CONCLUSION: Women elicited a greater response of serum GH and insulin to sprint exercise. This may contribute to explaining the earlier observed muscle hypertrophy in women in response to sprint training.

Inhibition of inducible nitric oxide synthase enhances anti-tumour immune responses in rats immunized with IFN-gamma-secreting glioma cells.

Interferon gamma (IFN-gamma) has successfully been used in immunotherapy of different experimental tumours. Mechanistically, IFN-gamma has extensive effects on the immune system including release of nitric oxide (NO) by upregulation of the inducible nitric oxide synthase (iNOS). NO has putative immunosuppressive effects but could also play a role in killing of tumour cells. Therefore, the aim of the present study was to clarify whether inhibition of iNOS in rats immunized with glioma cells (N32) producing IFN-gamma (N32-IFN-gamma), could enhance the anti-tumour immune response. Initially, both a selective iNOS, l-N(6)-(1-Iminoethyl)-l-lysine (l-NIL), and non-selective, N-nitro-l-arginine methyl ester (l-NAME), inhibitor of NOS were tested in vitro. After polyclonal stimulation with LPS and SEA, both l-NIL and l-NAME enhanced proliferation and production of IFN-gamma from activated rat splenocytes and this effect was inversely correlated to the production of NO. However, l-NIL had a broader window of efficacy and a lower minimal effective dose. When rats were immunized with N32-IFN-gamma, and administered NOS inhibitors by intraperitoneal (i.p.) mini-osmotic pumps, only splenocytes of rats treated with l-NIL, but not l-NAME, displayed an enhanced proliferation and production of IFN-gamma when re-stimulated with N32 tumour cells. Based on these findings, l-NIL was administered concurrently with N32-IFN-gamma cells to rats with intracerebral (i.c.) tumours resulting in a prolonged survival. These results show that inhibition of iNOS can enhance an IFN-gamma-based immunotherapy of experimental i.c. tumours implying that NO released after immunization has mainly immunosuppressive net effects.

Expression of oestrogen receptor alpha and beta is higher in skeletal muscle of highly endurance-trained than of moderately active men.

AIM: Two known oestrogen receptors (ERs), ERalpha and the recently cloned ERbeta, are expressed in the human skeletal muscle of both males and females. The effects of oestrogen and the role of ERs in skeletal muscle tissue are not well known. Oestrogen receptors and some of their target genes are involved in angiogenic processes. It was hypothesized that ERs are expressed at a higher level in a group with higher oxidative capacity, and that such an enhanced expression would parallel expression of the angiogenic factor -- vascular endothelial growth factor (VEGF). METHOD: Muscle biopsies were taken from vastus lateralis in 10 highly endurance-trained males and 10 moderately active males and analysed for the expression of ERs and VEGF. RESULTS: The major findings in the present study were the higher mRNA levels of ERalpha, ERbeta and VEGF in the highly endurance-trained than in the moderately active group. CONCLUSION: These data suggest that the greater mRNA expression of ERalpha and ERbeta and the oestrogen-associated angiogenic factor VEGF support the hypothesis of an involvement of ERs in the adaptation of skeletal muscle to endurance training.

Muscle fiber characteristics in female dancers during an active and an inactive period.

Muscle biopsies were taken from female dancers, aged 24 +/- 2 years, first during an active period and then after 32 weeks of detraining. During the active period, the percentage of type I fibers was 63% +/- 12%, and of the subgroups IIA, IIB, and IIC, 26% +/- 9%, 8% +/- 7%, and 3% +/- 3%, respectively. There was no significant change due to detraining either in the proportion of type I fibers or in the type II subgroups. During the active period, the cross-sectional area of the type I fibers was 3798 +/- 586 micron 2. The areas of the subgroups IIA, IIB, and IIC were 3394 +/- 858, 2497 +/- 530, and 3296 +/- 638 micron 2, respectively. After the detraining period, the area of the type I fibers had increased significantly (P less than 0.05). The areas of the types IIA, IIB, and IIC tended to be increased although not significantly. It is suggested that the comparatively high percentage of type I fibers, which was found in the active dancers, is the result of selection rather than of training, while the small fiber areas seem to be due to training.

Increase in the proportion of fast-twitch muscle fibres by sprint training in males.

Fifteen male physical education students were studied. The subjects trained for 4-6 weeks, 2-3 days per week, on a mechanically braked bicycle ergometer. A training session consisted of repeated 30-s 'all-out' sprints on a Wingate bicycle ergometer, on which the brake band of the flywheel was loaded with 75 g kg-1 body wt, with rest periods of 15-20 min between consecutive sprints. Thigh muscle biopsies were taken before and after the training period and were analysed for fibre types using a myofibrillar ATPase stain. The proportion of type I fibres decreased from 57 to 48% (P less than 0.05) and type IIA fibres increased from 32 to 38% (P less than 0.05). This study indicates that it is possible to achieve a fibre type transformation with high-intensity training. The effect of two-legged 'sprint' training on muscle fibre type composition may be related to a changed pattern of muscle fibre activation (e.g. an increased stimulation frequency). A change in fibre activation frequency may induce an increased synthesis of type II fibre myosin (fast myosin). Hormonal influences such as enhanced adrenergic stimulation of the muscle fibres cannot be excluded as a contributing factor, however.

Fast twitch fibres may predict anaerobic performance in both females and males.

The aim of the present study was to compare males and females with similar training backgrounds regarding the relationship between anaerobic performance and muscle characteristics and to test whether any of the analysed expressions of muscle characteristics could predict some of the difference in anaerobic performance between sexes. Subjects performed 30 s all-out sprints on a bicycle ergometer (Wingate test) and needle muscle biopsies were taken at rest. Peak and mean power were respectively 44% and 48% higher in males than in females. Activity of total lactate dehydrogenase (LD) was 33% higher and of M subunit of LD 38% higher in males. Anaerobic performance was directly related to the proportion of type II fibres, the relative M subunit activity or the activity of PFK in both males and females and the higher M subunit activity in males could predict some of the sex difference in anaerobic performance. It is suggested that anaerobic performance is directly related to fast contractile or/and anaerobic metabolic properties of skeletal muscle with no sex difference in this relationship. The difference in anaerobic performance between the sexes may partly be related to the sex difference in anaerobic metabolic properties of skeletal muscle.

Does chronic hypoxaemia induce transformations of fibre types?

The study comprised nine patients with chronic obstructive lung disease. Quadriceps muscle biopsies were studied with respect to fibre type composition before and after haemodilution that brought haemoglobin (Hb) to within normal limits. Ten days elapsed between the two biopsy occasions. The arterial oxygen tension (PaO2) and saturation (SaO2) were depressed to 8.4 +/- 2.0 kPa and 89 +/- 11% in the patients with chronic obstructive lung disease and increased to 9.2 +/- 2.1 and 91 +/- 8% with haemodilation. The type II fibre proportion was 71 +/- 12% before haemodilation and significantly higher than normal (reference group, see Aniansson et al. 1981). Following haemodilation the proportion of type II fibres decreased significantly to 60 +/- 14%. The proportion of type II fibres was directly related to the haemoglobin content before, but not after, haemodilation and was inversely related to PaO2 and SaO2 both before and after haemodilation. In conclusion, hypoxaemia may be a factor underlying the high proportion of type II fibres found in patients with chronic obstructive lung disease.

Effects of ischaemic training on force development and fibre-type composition in human skeletal muscle.

Force (peak torque) of m. quadriceps femoris was measured during 60 repeated, voluntary dynamic knee extensions in 10 men before and after a 4-week training regimen of one-legged cycle exercise. Biopsies for histochemical analysis were obtained from the lateral vastus muscle after the training period. One leg was trained with the blood flow to the leg muscles reduced by local supra-atmospheric external pressure of 50 mmHg ('Ischaemic leg, I-leg'). Employing the same work-load profile the other leg was trained at normal atmospheric pressure ('Non-restricted-flow leg, N-leg'). In response to I-training, Maximum Peak Torqued (MPT; the highest torque produced in any contraction) and Initial Peak Torque (IPT; the average peak torque of the initial 12 contractions) decreased by 8% (P less than 0.01) and 9% (P less than 0.001), respectively. Final Peak Torque (FPT; the average peak torque of the final 12 contractions) increased by 13% (P less than 0.05) after I-training. No changes in MPT, IPT or FPT occurred following N-training. After training the proportion of slow-twitch fibres was higher (P less than 0.05) and the mean slow-twitch fibre area was larger (P less than 0.05) in the I-than in the N-trained leg. The results indicate that blood flow-restricted training, in contrast to non-restricted-flow training, decreases maximum voluntary dynamic force, possibly by inducing an increase in the share of the muscle cross-sectional area consisting of slow-twitch fibres. That flow-restricted training improves maintenance of force during short-term local exercise may reflect ischaemically induced changes in the metabolic characteristics of skeletal muscle.

Effects of renal failure on skeletal muscle.

In this cross-sectional study, we examined biopsies from the vastus lateralis muscle of 13 predialytic uremic men (mean age 46 +/- 8 years). Their average glomerular filtration rate was 14 +/- 7 ml/min x 1.73 m2 and their maximal exercise capacity, measured by standardized exercise test on a bicycle ergometer, was 184 +/- 45 W (94% of the expected norm). The proportion of type I fibers (type I%) in the uremic group was similar to that of the reference group (42 +/- 11 vs. 41 +/- 8% NS). The proportion of type IIA fibers (type IIA%) in the uremic group was higher than in the reference group (44 +/- 10 compared to 35 +/- 9%, p < 0.05). The proportion of type IIB fibers (type IIB%) was lower than in the reference group (13 +/- 8 vs. 21 +/- 8%, p < 0.05). Type I fiber area was similar to that of the reference group (4,768 +/- 1,033 vs. 4,627 +/- 1,112 microns 2, NS). Type IIA and type IIB fiber areas tended to be smaller than those of the reference group (type IIA fiber area: 4,515 +/- 929 vs. 5,213 +/- 1,288 microns 2, NS; type IIB fiber area: 3,953 +/- 1,066 vs. 4,406 +/- 1,582 microns 2, NS) with a type IIA area/type I area ratio which was significantly lower than in the reference group. Citrate synthase activity was 0.48 +/- 0.08 mu kat/g in the uremic group and 0.50 +/- 0.08 mu kat/g in the reference group, NS.(ABSTRACT TRUNCATED AT 250 WORDS)

Muscle fibre types and enzyme activities after training with local leg ischaemia in man.

Eight healthy men performed supine one-legged training on a bicycle ergometer 45 min per leg four times per week for 4 week. The ergometer and lower body were inside a pressure chamber, the opening of which was sealed at the level of the crotch. One leg trained with impeded leg blood flow (I-leg), induced by an increased (50 mmHg) chamber pressure, at the highest tolerable intensity. The contralateral leg trained at the same power under normal pressure (N-leg). Before and after training biopsies were taken from the vastus lateralis of both legs and maximal one-legged exercise tests were executed with both legs. Biopsies were repeated when the subjects had been back to their habitual physical activity for 3 months. Training increased exercise time to exhaustion, but more in the I-leg than in the N-leg. After training, the I-leg had higher activity of citrate synthase (CS), a marker of oxidative capacity, and lower activity of the M-subunit of lactate dehydrogenase isoenzymes. It also had a higher percentage of type-I fibres and a lower percentage of IIB fibres, larger areas of all fibre types and a greater number of capillaries per fibre. It is concluded that ischaemic training changes the muscle metabolic profile in a direction facilitating aerobic metabolism. An altered fibre-type composition may contribute, but is not enough prerequisite for the change.

Whole-muscle and single-fibre contractile properties and myosin heavy chain isoforms in humans.

The contractile characteristics of three human muscle groups (triceps surae, quadriceps femoris and triceps brachii) of seven young male subjects were examined. The contractile properties were determined from electrically evoked isometric responses and compared with fibre type composition determined from needle biopsy samples. Fibre types were identified using myosin heavy chain (MHC) isoforms as molecular markers with gel electrophoresis (SDS-PAGE) and histochemical ATPase staining. Four contractile parameters (twitch time to peak torque, the maximal rate of torque development, frequency response and fatiguability) were found to be related to fibre type composition. From the biopsy samples, single muscle fibres were isolated and chemically skinned. Isometric tension (Po) unloaded shortening velocity (Vo) and rate of tension rise (dP/dt) were determined. Each fibre was classified on the basis of its MHC isoform composition determined by SDS-PAGE. Fibres belonging to the same type showed identical contractile parameters regardless of the muscle of origin, except minor differences in Po of the fast fibres and dP/dt of slow fibres. The results are in favour of the conclusion that fibre type composition, determined using MHC isoforms as markers, is the major determinant of the diversity of contractile properties among human muscle groups.