Cryo-electron microscopy analysis of myosin at work and at rest

Myosins are a superfamily of ATP-driven actin-dependent molecular motors that are responsible for diverse functions from muscle contraction to cell division. The resolution revolution in cryo-EM has enabled characterisation of the interaction of myosin with its actin track in several states of the myosin motor cycle, for multiple myosin classes, allowing increased insight into the force generation mechanism. A major advancement in our understanding of myosin-2 regulation has come through solving structures of its shutdown state, dysregulation of which is implicated in multiple diseases. This review will discuss what has been accomplished so far with cryoEM, what is still yet to do, but within reach, and how better understanding of myosin structure–function relationships may lead to future therapeutic interventions.     

Effect of adrenergic blockade on lymphocyte cytokine production at rest and during exercise

To examine the effect of exercise andadrenergic blockade on lymphocyte cytokine production, six men ingestedeither a placebo (control) or an - (prazosin hydrochloride) and-adrenoceptor antagonist (timolol malate) capsule (blockade, or BLK)2 h before performing 19 ± 1 min of supine bicycle exerciseat 78 ± 3% peak pulmonary uptake. Blood was collected before andafter exercise, stimulated with phorbol 12-myristate 13-acetate andionomycin, and surface stained for T (CD3⁺) and naturalkiller [NK (CD3CD56⁺)] lymphocyte surfaceantigens. Cells were permeabilized, stained for the intracellularcytokines interleukin (IL)-2 and interferon (IFN)-, and analyzedusing flow cytometry. BLK had no effect on the resting concentration ofstimulated cytokine-positive T and NK lymphocytes or the amount ofcytokine they were producing. Exercise resulted in an increase (P< 0.05) in the concentration of stimulated T and NK lymphocytesproducing cytokines in the circulation, but these cells produced less(P < 0.05) cytokine post- compared with preexercise.BLK attenuated (P < 0.05) the elevation in theconcentration of lymphocytes producing cytokines during exercise;however, BLK did not affect the amount of IL-2 and IFN- produced.These results suggest that adrenergic stimulation contributes to theexercise-induced increase in the concentration of lymphocytes in thecirculation; however, it does not appear to be responsible for theexercise-induced suppression in cytokine production.

     

The impact of aging and habitual physical activity on static respiratory work at rest and during exercise

We investigated the effects of aging on the elastic properties of lung tissue and the chest wall, simultaneously quantifying the contribution of each component to static inspiratory muscle work in resting and exercising adults. We further evaluated the interaction of aging and habitual physical activity on respiratory mechanics. Static lung volumes and elastic properties of the lung and chest wall (pressure-volume relaxation maneuvers) in 29 chronically sedentary and 29 habitually active subjects, grouped by age, were investigated: young (Y, 20-30 years), middle-aged (M, 40-50 years), and older (O, >60 years). Using static pressure-volume data, we computed the elastic work of breathing (joules per liter, J.l(-1)), including inspiratory muscle work, over resting and exercising tidal volume excursions. Elastic work of the lung (Y = 0.79 +/- 0.05; M = 0.47 +/- 0.05; O = 0.43 +/- 0.05 J.l(-1)) and chest wall (Y = -0.49 +/- 0.06; M = -0.12 +/- 0.07; O = 0.04 +/- 0.05 J.l(-1) ) changed significantly with age (P < 0.05). With aging, a parallel displacement of the chest wall pressure-volume curve resulted in a shift from energy being stored primarily during expiration to energy storage during inspiration, and driving expiration, both at rest and during exercise. Although deviating significantly from young adults, this did not significantly elevate static inspiratory muscle work but resulted in a redistribution of the tissues on which this work was performed and the phase of the respiratory cycle in which it occurred. Nevertheless, static inspiratory muscle work remained similar across age groups, at rest and during exercise, and habitual physical activity failed to influence these changes.     

Effects of a short-term strength training programme on lymphocyte subsets at rest in elderly humans

The effects of a short-term strength training programme on resting lymphocyte subsets and stress hormone concentrations were analysed in 32 elderly sedentary subjects. Out of these 32 subjects, 8 women and 8 men [mean age 70.1 (SEM 1.0) years] were randomly assigned to a 8-week strength training programme which consisted of three sets of eight repetitions at 80% of one repetition maximum, for leg press, bilateral leg extension and seated chest press, 3 days a week. The remaining 8 women and 8 men [mean age 70.5 (SEM 0.9) years] served as controls. Absolute counts of lymphocyte subsets (CD20+, CD3+, CD3+CD4+, CD3+CD8+, CD3-CD56+CD16+) were measured with a new technique combining fluorescent microspheres and flow cytometry. In the trained subjects, substantial increases in strength took place in one repetition maximum during the 8-week training period for leg press [from means of 20.7 (SEM 1.0) to 23.6 (SEM 1.0) N x kg(-1) LBM (lean body mass)], chest press [from means of 5.4 (SEM 0.3) to 6.2 (SEM 0.3) N x kg(-1) LBM] and bilateral leg extension [from means of 6.3 (SEM 0.2) to 7.4 (SEM 0.3) N x kg(-1) LBM] movements. Baseline cortisol concentration (P < 0.01), CD20+ cell count (P < 0.05), CD3+ cell count (P < 0.05), and CD4+ cell count (P < 0.01) decreased in both groups secondary to circannual variations between winter and summer. No significant effect of strength training on resting adrenaline, noradrenaline and cortisol concentrations or distributions of lymphocyte subsets at rest was observed. The main finding of this study was to demonstrate that 8-week is too short a duration for a strength training programme to modify counts of lymphocyte subsets at rest in elderly sedentary adults.     

The (human) respiratory rate at rest

Journal of Mathematical Biology - Biological tissues are composed of cells surrounded by the extracellular matrix (ECM). The ECM can be thought of as a fibrous polymer network, acting as a natural...     

The cortisol response to hypobaric hypoxia at rest and post-exercise

High altitude exposure normally leads to a marked natriuresis and diuresis. Acute mountain sickness is often associated with fluid retention, to which an elevated cortisol may contribute. Most investigators report a rise in resting cortisol with ascent, but little data exist regarding the cortisol response to a day trekking. We therefore measured salivary cortisol during ascent to > 5000 m in a cohort of between 42-45 subjects following a 6-h trek (samples taken between 15:30-16:30 h) and between 15-20 subjects at rest (morning samples taken between 08:00-09:00 h). Morning resting cortisol [nmol/l, mean±sd, (range)] was 5.5±2.9 (2.13-13.61) at 1300 m; 4.7±6.8 (1.4-27.02) at 3400 m, and significantly (p=0.002) rose between 4270 m [3.5±2.1 (1.4-8.34)] and 5150 m [14.5±30.3 (1.9-123.1)]. Post-exercise cortisol [nmol/l, mean±sd, (range)] dropped between 3400 m [7±6 (1.5-33.3)] and 4270 m [4.2±4.8 (1.4-29.5)] (p=0.001) followed by a significant rise in post-exercise cortisol between 4270 m [4.2±4.8 (1.4-29.5)] and 5 150 m [9.2±10.2 (1.4-61.3)] (p<0.001). There were no significant associations between severity of acute mountain sickness and cortisol levels. There was a significant though weak correlation between cortisol post-exercise at 5150 m and oxygen saturation at 5150 m (rho= - 0.451, p=0.004). In conclusion, this is the largest cohort to have their resting and post-exercise cortisol levels ascertained at high altitude. We confirm the previous findings of an elevated resting morning cortisol at > 5000 m, but present the novel finding that the cortisol response to a day trekking at HA appears suppressed at 4270 m.     

Fatigue response of rat medial longissimus muscles induced with electrical stimulation at various work/rest ratios

Muscle fatigue may be a precursor to workplace musculoskeletal disorders, with the low back resulting in the most frequently injured body part. Work/rest ratios have an effect on fatigue due to the amount of rest allowance provided following muscle contraction. This study explored various work/rest ratios by electrically stimulating rat medial longissimus muscles. A 3 V stimulus with 0.2 ms pulse duration was applied at a frequency of 30 Hz. There were four stimulation groups consisting of the following duty cycles (DC) and cycle times (CT): DC25%:CT20s, DC25%:CT280s, DC75%:CT20s, and DC75%:CT180s. Muscle fatigue was measured as a decrease in M-wave amplitude and area, and an increase in M-wave duration. The results indicated that fatigue occurred immediately in each of the groups. The higher duty cycle and shorter cycle time group resulted in significantly greater fatigue than the lower duty cycle and longer cycle time group, as measured by increased M-wave amplitude and area. A longer M-wave duration was observed in the high duty cycle long cycle time group. This suggests that the combination of low duty cycle and long cycle times leads to less fatigue. In high duty cycle scenarios, short cycle times result in less fatigue.     

Effect of exercise to rest ratio on plasma lactate concentration at work rates above and below maximum oxygen uptake

The metabolic and physiological responses to different exercise to rest ratios (E:R) (2:1, 1:1, 1:2) of eight subjects exercising at work rates approximately 10% above and below maximum oxygen uptake (VO2max) were assessed. Each of the six protocols consisted of 15 1-min-long E:R intervals. Total work (kJ), oxygen uptake (VO2), heart rate (fc) and plasma lactate concentrations were monitored. With increases in either E:R or work rate, VO2 and fc increased (P < 0.05). The average (15 min) VO2 and fc ranged from 40 to 81%, and from 62 to 91% of maximum, respectively. Plasma lactate concentrations nearly doubled at each E:R when work rate was increased from 90 to 110% of VO2max and ranged from a low of 1.8 mmol.l-1 (1:2-90) to a high of 10.7 mmol.l-1 (2:1-110). The 2:1-110 protocol elicited plasma lactate concentrations which were approximately 15 times greater than that of rest. These data suggest that plasma lactate concentrations during intermittent exercise are very sensitive to both work rate and exercise duration.