The individual and combined effects of obesity- and ageing-induced systemic inflammation on human skeletal muscle properties.

BACKGROUND/OBJECTIVES: The purpose of this study was to determine whether circulating pro-inflammatory cytokines, elevated with increased fat mass and ageing, were associated with muscle properties in young and older people with variable adiposity. SUBJECTS/METHODS: Seventy-five young (18-49 yrs) and 67 older (50-80 yrs) healthy, untrained men and women (BMI: 17-49 kg/m2) performed isometric and isokinetic plantar flexor maximum voluntary contractions (MVCs). Volume (Vm), fascicle pennation angle (FPA), and physiological cross-sectional area (PCSA) of the gastrocnemius medialis (GM) muscle were measured using ultrasonography. Voluntary muscle activation (VA) was assessed using electrical stimulation. GM specific force was calculated as GM fascicle force/PCSA. Percentage body fat (BF%), body fat mass (BFM), and lean mass (BLM) were assessed using dual-energy X-ray absorptiometry. Serum concentration of 12 cytokines was measured using multiplex luminometry. RESULTS: Despite greater Vm, FPA, and PCSA (P<0.05), young individuals with BF% ⩾40 exhibited 37% less GM specific force compared to young BF%<40 (P<0.05). Older adults with BF% ⩾40 showed greater isokinetic MVC compared to older BF%<40 (P=0.019) but this was reversed when normalised to body mass (P<0.001). IL-6 correlated inversely with VA in young (r=-0.376; P=0.022) but not older adults (p>0.05), while IL-8 correlated with VA in older but not young adults (r⩾0.378, P⩽0.027). TNF-alpha correlated with MVC, lean mass, GM FPA and maximum force in older adults (r⩾0.458; P⩽0.048). CONCLUSIONS: The age- and adiposity-dependent relationships found here provide evidence that circulating pro-inflammatory cytokines may play different roles in muscle remodelling according to the age and adiposity of the individual.

Omega-3 Fatty Acids and Vitamin D in Immobilisation: Part A- Modulation of Appendicular Mass Content, Composition and Structure.

OBJECTIVES: Muscle size decreases in response to short-term limb immobilisation. This study set out to determine whether two potential protein-sparing modulators (eicosapentaenoic acid and vitamin D) would attenuate immobilisation-induced changes in muscle characteristics. DESIGN: The study used a randomised, double-blind, placebo-controlled design. SETTING: The study took part in a laboratory setting. PARTICIPANTS: Twenty-four male and female healthy participants, aged 23.0±5.8 years. INTERVENTION: The non-dominant arm was immobilised in a sling for a period of nine waking hours a day over two continuous weeks. Participants were randomly assigned to one of three groups: placebo (n=8, Lecithin, 2400 mg daily), omega-3 (ω-3) fatty acids (n=8, eicosapentaenoic acid (EPA); 1770 mg, and docosahexaenoic acid (DHA); 390 mg, daily) or vitamin D (n=8, 1,000 IU daily). MEASUREMENTS: Muscle and sub-cutaneous adipose thickness (B-mode ultrasonography), body composition (DXA) and arm girth (anthropometry) were measured before immobilisation, immediately on removal of the sling and two weeks after re-mobilisation. RESULTS: Muscle thickness (-5.4±4.3%), upper and lower arm girth (-1.3±0.4 and -0.8±0.8%, respectively), lean mass (-3.6±3.7%) and bone mineral content (BMC) (-2.3±1.5%) decreased significantly with limb immobilisation in the placebo group (P<0.05). Despite no significant effect of group, ω-3 and vitamin D supplementation showed trends (p>0.05) towards attenuating the decreases in muscle thickness, upper/lower arm girths and BMC observed in the placebo group. The ω-3 supplementation group demonstrated a non-significant attenuation of the decrease in DXA quantified lean mass observed in the placebo group. Sub-cutaneous adipose thickness increased in the placebo group (P<0.05). ω-3 and vitamin D both blunted this response, with ω-3 having a greater effect (P<0.05). All parameters had returned to baseline values at the re-mobilisation phase of the study. CONCLUSION: Overall, at the current doses, ω-3 and vitamin D supplementation only attenuated one of the changes associated with non-injurious limb immobilisation. These findings would necessitate further research into either a) supplementation linked to injury-induced immobilisation, or b) larger doses of these supplements to confirm/refute the physiological reserve potential of the two supplements.

Omega-3 Fatty Acids and Vitamin D in Immobilisation: Part B- Modulation of Muscle Functional, Vascular and Activation Profiles.

OBJECTIVES: This study set out to determine whether two potential protein-sparing modulators (eicosapentaenoic acid and vitamin D) would modulate the anticipated muscle functional and related blood vessels function deleterious effects of immobilisation. DESIGN: The study used a randomised, double-blind, placebo-controlled design. SETTING: The study took part in a laboratory setting. PARTICIPANTS: Twenty-four male and female healthy participants, aged 23.0±5.8 years. INTERVENTION: The non-dominant arm was immobilised in a sling for a period of nine waking hours a day over two continuous weeks. Participants were randomly assigned to one of three groups: placebo (n=8, Lecithin, 2400 mg daily), omega-3 (ω-3) fatty acids (n=8, eicosapentaenoic acid (EPA); 1770 mg, and docosahexaenoic acid (DHA); 390 mg, daily) or vitamin D (n=8, 1,000 IU daily). MEASUREMENTS: Isometric and isokinetic torque, antagonist muscle co-contraction (activation profile), muscle fatigability indices, and arterial resting blood flow were measured before, at the end of the immobilisation period, and two weeks after re-mobilisation. RESULTS: Muscle elbow flexion and extension isometric and isokinetic torque decreased significantly with limb immobilisation in the placebo group (P<0.05). Despite no significant effect of supplementation, ω-3 and vitamin D supplementation showed trends (P>0.05) towards attenuating the decreases observed in the placebo group. There was no significant change in muscle fatigue parameters or co-contraction values with immobilisation and no effect of supplementation group (P>0.05). Similarly, this immobilisation model had no impact on the assessed blood flow characteristics. All parameters had returned to baseline values at the re-mobilisation phase of the study. CONCLUSION: Overall, at the current doses, neither ω-3 nor vitamin D supplementation significantly attenuated declines in torque associated with immobilisation. It would appear that muscle function (described here in Part B) might not be as useful a marker of the effectiveness of a supplement against the impact of immobilisation compared to tissue composition changes (described in Part A).

Muscle Damage following Maximal Eccentric Knee Extensions in Males and Females.

AIM: To investigate whether there is a sex difference in exercise induced muscle damage. MATERIALS AND METHOD: Vastus Lateralis and patella tendon properties were measured in males and females using ultrasonography. During maximal voluntary eccentric knee extensions (12 reps x 6 sets), Vastus Lateralis fascicle lengthening and maximal voluntary eccentric knee extensions torque were recorded every 10° of knee joint angle (20-90°). Isometric torque, Creatine Kinase and muscle soreness were measured pre, post, 48, 96 and 168 hours post damage as markers of exercise induced muscle damage. RESULTS: Patella tendon stiffness and Vastus Lateralis fascicle lengthening were significantly higher in males compared to females (p<0.05). There was no sex difference in isometric torque loss and muscle soreness post exercise induced muscle damage (p>0.05). Creatine Kinase levels post exercise induced muscle damage were higher in males compared to females (p<0.05), and remained higher when maximal voluntary eccentric knee extension torque, relative to estimated quadriceps anatomical cross sectional area, was taken as a covariate (p<0.05). CONCLUSION: Based on isometric torque loss, there is no sex difference in exercise induced muscle damage. The higher Creatine Kinase in males could not be explained by differences in maximal voluntary eccentric knee extension torque, Vastus Lateralis fascicle lengthening and patella tendon stiffness. Further research is required to understand the significant sex differences in Creatine Kinase levels following exercise induced muscle damage.

The emergence of sedentary behaviour physiology and its effects on the cardiometabolic profile in young and older adults.

It has recently emerged that sedentary behaviour is independent of a lack of physical activity as individuals can be sufficiently active, based on the recommended physical activity guidelines, but also spend the majority of their waking hours engaging in sedentary behaviour. Individuals who follow this pattern of physical activity and sedentary behaviour are known as 'active couch potatoes'. Sedentary behaviour has been found to have detrimental effects on cardiometabolic markers associated with cardiovascular disease. Since the positive effects of moderate-to-vigorous intensity physical activity do not necessarily negate the deleterious effects of sedentary behaviour on cardiometabolic markers, it is postulated that engaging in light physical activity is an intervention that will successfully reduce levels of sedentary behaviour and may hence improve health markers of quality of life. We propose that such lifestyle changes may be particularly relevant to older populations as these engage in sedentary behaviour for the majority of their waking hours, thereby adding to the negative aging effect on cardiometabolic markers.

The impact of obesity on skeletal muscle architecture in untrained young vs. old women.

It is unknown whether loading of the lower limbs through additional storage of fat mass as evident in obesity would promote muscular adaptations similar to those seen with resistance exercise. It is also unclear whether ageing modulates any such adjustments. This study aimed to examine the relationships between adiposity, ageing and skeletal muscle size and architecture. A total of 100 untrained healthy women were categorised by age into young (Y) (mean ± SD: 26.7 ± 9.4 years) vs. old (O) (65.1 ± 7.2 years) and body mass index (BMI) classification (underweight, normal weight, overweight and obese). Participants were assessed for body fat using dual energy x-ray absorptiometry, and for gastrocnemius medialis (GM) muscle architecture (skeletal muscle fascicle pennation angle and length) and size [GM muscle volume and physiological cross-sectional area (PCSA)] using B-mode ultrasonography. GM fascicle pennation angle (FPA) in the obese Y females was 25% greater than underweight (P = 0.001) and 25% greater than normal weight (P = 0.001) individuals, while O females had 32 and 22% greater FPA than their underweight (P = 0.008) and normal weight (P = 0.003) counterparts. Furthermore, FPA correlated with body mass in both Y and O females (Y r = 0.303; P < 0.001; O r = 0.223; P = 0.001), yet no age-related differences in the slope or r-values were observed (P > 0.05). Both GM muscle volume (P = 0.003) and PCSA (P = 0.004) exhibited significant age × BMI interactions. In addition, muscle volume and PCSA correlated with BMI, body mass and fat mass. Interestingly, ageing reduced both the degree of association in these correlations (P < 0.05) and the slope of the regressions (P < 0.05). Our findings partly support our hypotheses in that obesity-associated changes in GM PCSA and volume differed between the young and old. The younger GM muscle adapted to the loading induced by high levels of body mass, adiposity and BMI by increasing its volume and increasing its pennation angle, ultimately enabling it to produce higher maximum torque. Such an adaptation to increased loading did not occur in the older GM muscle. Nonetheless, the older GM muscle FPA increased to a similar extent to that seen in young GM muscle, an effect which partly explains the relatively enhanced absolute maximum torque observed in obese older females.

Passive stiffness of the gastrocnemius muscle in athletes with spastic hemiplegic cerebral palsy.

The passive properties of the muscle-tendon unit are regularly assessed in individuals with cerebral palsy (CP). However, no information is available on the passive properties of adult muscle, and whether any differences exist between the paretic and control muscles. Eleven ambulant male athletes with spastic hemiplegic CP (21.2 ± 3.0 years) and controls without neurological impairment (age = 21.8 ± 2.2 years) completed two and one passive stretch session, respectively. During each session, the ankle was passively dorsiflexed until end range of motion (ROM), whilst recording passive ankle angle, torque and gastrocnemius medialis (GM) myotendinous junction (MTJ) displacement. In addition, GM cross-sectional area (CSA) and length were measured. Subsequently, in vivo stress and strain were determined to calculate elastic modulus. Passive stiffness, MTJ displacement and ROM of the paretic GM were not different from the control muscles. However, the elastic modulus of the paretic GM was two times stiffer than the control GM muscles. In conclusion, athletes with CP exhibit absolute passive muscle stiffness similar to the controls; however, the elastic modulus of the CP muscle was significantly greater. Therefore, throughout the same ROM a smaller GM CSA in CP athletes has to dissipate larger relative torque compared to the control muscles, consequently causing the muscle to elongate to the same extent as the non-paretic muscle under stretch.

The effect of the oral contraceptive pill on the passive stiffness of the human gastrocnemius muscle in vivo.

The aim of this investigation was to determine the effect of sustained monophasic oral contraceptive pill (MOCP) use on the in vivo passive stiffness of the gastrocnemius medialis (GM) muscle-tendon unit. Twenty four females volunteered for this study (age range 20-25 yrs); twelve participants had been taking the combined MOCP for a minimum of 12 months, and twelve participants, who had never taken the MOCP, formed a control group. Distal displacement of the GM myotendinous junction (MTJ) was measured during passive dorsiflexion at 2 Nm increments to 20 Nm, and at end range of motion using ultrasonography. In addition, GM MTJ displacement was measured at passive torques equivalent to 5, 10 and 15% of plantarflexion maximal voluntary contraction (MVC) torque, and relative to GM length. MOCP users had significantly greater GM MTJ displacement at all passive torques (P<0.01), reaching 40% more at 20 Nm; these displacements remained significantly different when MVC and GM length were accounted for (P<0.01). Passive muscle stiffness from 0-20 Nm was 31% less in MOCP users compared to non-users (P<0.01). In conclusion, based on the in vivo assessment of GM MTJ displacement, passive muscle stiffness is less in MOCP using females, compared to non-pill users.

Serum relaxin levels affect the in vivo properties of some but not all tendons in normally menstruating young women.

Relaxin (hRLX) is a hormone reported to affect collagen synthesis. Its effects are also thought to be modulated by other sex hormones, including oestrogen, which has previously been found to be associated with alterations of in vivo tendon properties. There is thus a potential for hRLX to impact on collagen, which could result in tendon structural and mechanical properties being modified. The present study therefore aimed to determine any interaction between hRLX and tendon stiffness, in normally menstruating women (n = 12). Tendon properties were determined using a combination of dynamometry and B-mode ultrasound, whilst serum hRLX levels were established by ELISA. Serum hRLX level was seen to be negatively associated with patellar tendon stiffness (r = -0.56; P < 0.001), explaining 31% of the variance in this parameter. There was no association between hRLX and gastrocnemius tendon stiffness (P > 0.05), or with the cross-sectional area of either of the two tendons (P > 0.05). In young, normally menstruating women, hRLX appears to have a significant effect on the patellar but not the gastrocnemius tendon stiffness. Where it has an effect, this appears to be on the intrinsic properties rather than on the dimensions of said tendon. Future work to elucidate the physiological cause of this selectivity in the impact of relaxin will be key to mapping the impact of the endocrine system on the phenotype of tendinous tissue.

Menstrual cycle variations in oestradiol and progesterone have no impact on in vivo medial gastrocnemius tendon mechanical properties.

BACKGROUND: Tendon tissue contains oestrogen receptors and is therefore likely to be responsive to female sex hormones. Here we examine any effect of levels of female sex hormones associated with the menstrual cycle phase on corresponding tendon mechanical properties. METHODS: Fifteen healthy females aged 23 (SEM 1.0 years) underwent three assessments of medial gastrocnemius tendon mechanical properties. Assessments were carried out once during days 1-4, 12-14 and 20-23 (with day 1 being the first day of menstruation). Venous blood samples were taken on the same days as tendon properties assessments to quantify serum levels of oestradiol and progesterone. FINDINGS: There was no significant difference in the stiffness of the medial gastrocnemius tendon over the course of the menstrual cycle (days 1-4, 65.08 (SEM 5.16 Nm m(-1)), days 12-14, 62.73 (SEM 5.82 Nm m(-1)), days 20-23, 66.74 (SEM 7.14 Nm m(-1))). There were also no significant differences in tendon length and cross-sectional area which led to no significant differences in Young's modulus values. No correlations were found between serum levels of oestradiol and/or progesterone and tendon stiffness and/or Young's modulus. INTERPRETATION: Acute fluctuations in female sex hormones have no significant effect on medial gastrocnemius tendon mechanical properties. In a context where studies are often limited to selecting only oral contraceptive-users as participants in order to minimise potential noise related to the anticipated effects of menstrual cycle hormones on physical performance, our findings provide the basis for enabling the pooling of female tendon data, regardless of the phase of the menstrual cycle of individual participant.

Tendon structural and mechanical properties do not differ between genders in a healthy community-dwelling elderly population.

Elderly women are reportedly at higher risk of falling than their male counterparts. Postural balance is highly associated with fall risk and is also correlated with tendon structural and mechanical properties. Gender differences in tendon properties could partly explain the discrepancy in fall risk. Thus the purpose of this study was to investigate the possible gender difference in tendon properties in the elderly. The properties of the patellar tendon of 55 elderly (men n = 27, aged 72 +/- 1 years, women n = 28, aged 70 +/- 1 years) participants were tested. Tendon stiffness (K), length (L), and cross-sectional area (CSA) were measured using B-mode ultrasonography, dynamometry, and electromyography during ramped isometric knee extensions. There were no significant differences (p > 0.05) between men and women in tendon stiffness (elderly men 550.9 +/- 29.2 vs. women 502.9 +/- 44.9 Nmm(-1)) or in Young's modulus (elderly men 0.32 +/- 0.02 vs. women 0.36 +/- 0.04 GPa). This elderly group had similar tendon structural and mechanical properties. The comparable characteristics in gender-specific tendon properties in an elderly population exhibiting similar lifestyle characteristics to the current sample may not explain the reports in the literature regarding increased fall risk in elderly women relative to that seen in men of a similar age.

Improvements in muscle-tendon properties are beneficial to balance in multiple sclerosis.

A 63-year-old male was diagnosed with multiple sclerosis (MS) two years prior to this investigation. We hypothesized that compromised postural balance in MS can be improved by training-induced alterations in muscle-tendon properties. Sixteen weeks of strength and balance training induced an increase in quadriceps and gastrocnemius muscle size, strength, central activation, muscle recruitment and tendon stiffness. Concomitantly, the ability to maintain postural stances was increased in both eyes-open and eyes-closed conditions. We concluded that improvements in muscle-tendon properties were accompanied by improved balance and a general feeling of well-being after training.

Oestrogen status in relation to the early training responses in human thumb adductor muscles.

AIMS: The aims of this study were to identify the mechanisms for the early response to training in women of different oestrogen status and to determine whether any oestrogen and exercise effects on these would be additive. METHODS: We monitored training (ten 5-s contractions per day for 12 weeks)-induced changes in the size, strength, voluntary activation capacity and index of crossbridge force state (i.e. rapid stretch to isometric torque ratio), in the thumb adductor muscles of postmenopausal [eight who had never used, and 14 who were using, hormone replacement therapy (HRT)] and seven premenopausal eumenorrhoeic women. The contralateral untrained muscle was used as a control. RESULTS: There was a significant effect of oestrogen status on the magnitude of training-induced strength increment, with the non-HRT postmenopausal group exhibiting the greatest benefits (28 +/- 6%, P = 0.024) from training. There were no significant or commensurate changes in either cross-sectional area or voluntary activation capacity. The index of crossbridge force state improved most in the no-HRT group (19 +/- 7%, P < 0.05). CONCLUSIONS: Presence, rather than absence of oestrogen, is associated with relatively higher muscle function which limits the potential for any further training-induced increments in muscle performance, as would be expected if the muscle strengthening actions of training and oestrogen share a common, partially saturable physiological pathway. The mechanism that is involved in the early training-induced strength increment in the three differing oestrogen groups cannot be due to increased size or recruitment. It would appear instead that increased motor unit firing frequency is involved.

IL-6-174G/C genotype is associated with the bone mineral density response to oestrogen replacement therapy in post-menopausal women.

A reduction in interleukin-6 (IL-6) activity may contribute to the beneficial effects of hormone replacement therapy (HRT) on the menopausal decline in bone mineral density (BMD). We have examined this hypothesis using a genetic strategy. The -174C (rather than G) IL-6 gene variant is associated with lower IL-6 expression. As such, we might anticipate the C allele to be associated with a greater response to HRT. We have tested this hypothesis. Mean three-site [spine (L1-L4), neck of femur, and Ward's triangle] BMD was measured in 65 women in a 1-year randomised controlled trial of HRT with 0.625 mg oestrogen/day and 0.15 mg norgestrel (n=30). Baseline BMD was genotype-independent for both the control and HRT group. In the control group, the percentage change in BMD after 1 year was similar between genotypes (P=0.45). In contrast, in the HRT group, the rise was genotype-dependent. Those homozygous for the G allele showed a 3.62 (2.14)% increase in BMD compared with 10.44 (4.68)% for the C-homozygous group. Heterozygotes had an intermediate BMD increase of 5.6 (2.82)% [ P=0.006 ( P value for interaction between HRT and genotype was 0.04)] Although the study was limited by its small sample size, these are the first data to demonstrate the importance of IL-6 genotype in determining response to oestrogen therapy, rather than its physiological withdrawal.

Angiotensin-I converting enzyme genotype-dependent benefit from hormone replacement therapy in isometric muscle strength and bone mineral density.

Low bone mineral density (BMD) and muscle weakness are major risk factors for postmenopausal osteoporotic fracture. Hormone replacement therapy (HRT) reverses the menopausal decline in maximum voluntary force of the adductor pollicis and reduces serum angiotensin-I converting enzyme (ACE) levels. The insertion (I) allele of the ACE gene polymorphism is associated with lower ACE activity and improved muscle efficiency in response to physical training. Therefore, we examined whether the presence of the I allele in postmenopausal women would affect the muscle response to HRT. Those taking HRT showed a significant gain in normalized muscle maximum voluntary force slope, the rate of which was strongly influenced by ACE genotype (16.0 +/- 1.53%, 14.3 +/- 2.67%, and 7.76 +/- 4.13%, mean +/- SEM for II, ID, and DD genotype, respectively; P = 0.017 for gene effect, P = 0.004 for I allele effect). There was also a significant ACE gene effect in the response of BMD to HRT in Ward's triangle (P = 0.03) and a significant I allele effect in the spine (P = 0.03), but not in the neck of femur or total hip. These data suggests that low ACE activity associated with the I allele confers an improved muscle and BMD response in postmenopausal women treated with HRT.