Biomedical subjects

M Hargreaves

Publications and source records attributed to M Hargreaves.

At least 37 records · Page 2Linked to original sources

PubMed2000

Fiber type-specific expression of GLUT4 in human skeletal muscle: influence of exercise training.

The fiber type-specific expression of skeletal muscle GLUT4 and the effect of 2 weeks of low-intensity training were investigated in 8 young untrained male subjects. Single muscle fibers were dissected from a vastus lateralis biopsy sample. Based on myosin heavy chain (MHC) expression, fibers were pooled into 3 groups (MHC I, MHC IIA, and MHC IIX), and the GLUT4 content of 15-40 pooled fibers was determined using SDS-PAGE and immunological detection. The GLUT4 content in pooled muscle fibers expressing MHC I was approximately 20% higher (P < 0.05) than that in muscle fibers expressing MHC IIA or MHC IIX. No difference in GLUT4 could be detected between fibers expressing MHC IIA or MHC IIX. Two weeks of exercise training increased (P < 0.05) the peak power output of the knee extensors by 13%, the maximal activities of citrate synthase and 3-hydroxyacyl-CoA dehydrogenase by 21 and 18%, respectively, and the GLUT4 protein content by 26% in a muscle homogenate. Furthermore, a 23% increase (P < 0.05) in GLUT4 was seen in fibers expressing the MHC I isoform after exercise training for 2 weeks. No change was seen in fibers expressing MHC IIA or MHC IIX. In conclusion, our data directly demonstrate that GLUT4 is expressed in a fiber type-specific manner in human skeletal muscle, although fiber type differences are relatively small. In addition, low-intensity exercise training recruiting primarily fibers expressing MHC I increased GLUT4 content in these fibers but not in fibers expressing MHC IIA or MHC IIX, indicating that GLUT4 protein content is related more to activity level of the fiber than to its fiber type, which is defined by expression of contractile protein.

Adult↗

PubMed1999

Glucose production during strenuous exercise in humans: role of epinephrine.

The increase in hepatic glucose production (HGP) that occurs during intense exercise is accompanied by a simultaneous increase in epinephrine, which suggests that epinephrine may be important in regulating HGP. To further investigate this, six trained men were studied twice. The first trial [control (Con)] consisted of 20 min of cycling at 40 +/- 1% peak oxygen uptake (VO2 peak) followed by 20 min at 80 +/- 2% VO2 peak. During the second trial [epinephrine (Epi)], subjects exercised for 40 min at 41 +/- 2% VO2 peak. Epinephrine was infused during the latter 20 min of exercise and resulted in plasma levels similar to those measured during intense exercise in Con. Glucose kinetics were measured using a primed, continuous infusion of [3-3H]glucose. HGP was similar at rest (Con, 11.0 +/- 0.5 and Epi, 11.1 +/- 0.5 micromol. kg-1. min-1). In Con, HGP increased (P < 0.05) during exercise to 41.0 +/- 5.2 micromol. kg-1. min-1 at 40 min. In Epi, HGP was similar to Con during the first 20 min of exercise. Epinephrine infusion increased (P < 0.05) HGP to 24.0 +/- 2.5 micromol. kg-1. min-1 at 40 min, although this was less (P < 0.05) than the value in Con. The results suggest that epinephrine can increase HGP during exercise in trained men; however, epinephrine during intense exercise cannot fully account for the rise in HGP. Other glucoregulatory factors must contribute to the increase in HGP during intense exercise.

Adult↗

PubMed1999

Effect of short-term training on mitochondrial ATP production rate in human skeletal muscle.

Seven untrained volunteers [3 men, 4 women, 20.1 +/- 2.0 (SD) yr, 66. 0 +/- 11.0 kg, 171 +/- 13 cm] participated in a 10-day cycle exercise training program. Resting muscle samples were obtained from vastus lateralis before and after 5 and 10 days of training. Mitochondrial ATP production rate (MAPR) was assayed in isolated mitochondria by using a bioluminescence technique and referenced to the activity of glutamate dehydrogenase in the muscle sample. MAPR increased 136 and 161% after 10 days of training for the mitochondrial substrate combinations pyruvate + palmitoyl-L-carnitine + alpha-ketoglutarate + malate and palmitoyl-L-carnitine + malate, respectively. Total muscle glutamate dehydrogenase and citrate synthase activity increased 53 and 16%, respectively, after 5 days but did not significantly increase further after 10 days. The results from the present study indicate that MAPR, measured by using the substrate combinations pyruvate + palmitoyl-L-carnitine + alpha-ketoglutarate + malate and palmitoyl-L-carnitine + malate, can rapidly increase in response to endurance training.

Adenosine Triphosphate↗

PubMed1999

Muscle metabolism during prolonged exercise in humans: influence of carbohydrate availability.

Eight endurance-trained men cycled to volitional exhaustion at 69 +/- 1% peak oxygen uptake on two occasions to examine the effect of carbohydrate supplementation during exercise on muscle energy metabolism. Subjects ingested an 8% carbohydrate solution (CHO trial) or a sweet placebo (Con trial) in a double-blind, randomized order, with vastus lateralis muscle biopsies (n = 7) obtained before and immediately after exercise. No differences in oxygen uptake, heart rate, or respiratory exchange ratio during exercise were observed between the trials. Exercise time to exhaustion was increased by approximately 30% when carbohydrate was ingested [199 +/- 21 vs. 152 +/- 9 (SE) min, P < 0.05]. Plasma glucose and insulin levels during exercise were higher and plasma free fatty acids lower in the CHO trial. No differences between trials were observed in the decreases in muscle glycogen and phosphocreatine or the increases in muscle lactate due to exercise. Muscle ATP levels were not altered by exercise in either trial. There was a small but significant increase in muscle inosine monophosphate levels at the point of exhaustion in both trials, and despite the subjects in CHO trial cycling 47 min longer, their muscle inosine monophosphate level was significantly lower than in the Con trial (CHO: 0.16 +/- 0.08, Con: 0.23 +/- 0.09 mmol/kg dry muscle). These data suggest that carbohydrate ingestion may increase endurance capacity, at least in part, by improving muscle energy balance.

Adult↗

PubMed1999

Acute plasma volume expansion: effect on metabolism during submaximal exercise.

To examine the effect of acute plasma volume expansion (PVE) on substrate selection during exercise, seven untrained men cycled for 40 min at 72 +/- 2% peak oxygen uptake (VO(2 peak)) on two occasions. On one occasion, subjects had their plasma volume expanded by 12 +/- 2% via an intravenous infusion of the plasma substitute Haemaccel, whereas on the other occasion no such infusion took place. Muscle samples were obtained before and immediately after exercise. In addition, heart rate and pulmonary gas and venous blood samples were obtained throughout exercise. No differences in oxygen uptake or heart rate during exercise were observed between trials, whereas respiratory exchange ratio, blood glucose, and lactate were unaffected by PVE. Muscle glycogen and lactate concentrations were not different either before or after exercise. In addition, there was no difference in total carbohydrate oxidation between trials (control: 108 +/- 2 g; PVE group: 105 +/- 2 g). Plasma catecholamine levels were not affected by PVE. These data indicate that substrate metabolism during submaximal exercise in untrained men is unaltered by acute hypervolemia.

Adult↗

PubMed1999

Effect of temperature on muscle metabolism during submaximal exercise in humans.

To study the effect of temperature on muscle metabolism during submaximal exercise, six endurance-trained men had one thigh warmed and the other cooled for 40 min prior to exercise using water-perfused cuffs. One cuff was perfused with water at 50-55 degrees C (HL) with the other being perfused with water at 0 degree C (CL). With the cuffs still in position, subjects performed cycling exercise for 20 min at a work load corresponding to 70% VO2,peak (where VO2,peak is peak pulmonary oxygen uptake) in comfortable ambient conditions (20-22 degrees C). Muscle biopsies were obtained prior to and following exercise and forearm venous blood was collected prior to and throughout the exercise period. Muscle temperature (Tmus) was not different prior to treatment, but treatment resulted in a large difference in pre-exercise Tmus (difference = 6.9 +/- 0.9 degrees C; P < 0.01). Although this difference was reduced following exercise; it was nonetheless significant (difference = 0.4 +/- 0.1 degree C; P < 0.05). Intramuscular [ATP] was not affected by either exercise or muscle temperature. [Phosphocreatine] decreased (P < 0.01) and [creatine] increased (P < 0.01) with exercise but were not different when comparing HL with CL. Muscle lactate concentration was not different prior to treatment nor following exercise when comparing HL with CL. Muscle glycogen concentration was not different when comparing the trials before treatment, but the post-exercise value was lower (P < 0.05) in HL compared with CL. Thus, net muscle glycogen use was greater during exercise with heating (208 +/- 23 vs. 118 +/- 22 mmol kg-1 for HL and CL, respectively; P < 0.05). These data demonstrate that muscle glycogen use is augmented by increases in intramuscular temperature despite no differences in high energy phosphagen metabolism being observed when comparing treatments. This suggests that the increase in carbohydrate utilization occurred as a direct effect of an elevated muscle temperature and was not secondary to allosteric activation of enzymes mediated by a reduced ATP content.

Adenosine Triphosphate↗

PubMed1998

1997 Sir William Refshauge Lecture. Skeletal muscle glucose metabolism during exercise: implications for health and performance.

Skeletal muscle glucose uptake and metabolism are major determinants of whole body glucose metabolism in response to exercise and insulin stimulation. An understanding of the mechanisms responsible for increased muscle glucose uptake under these conditions is crucial for identifying strategies that enhance insulin action and exercise performance. Regular exercise, by favourably influencing the intramuscular determinants of glucose uptake, enhances insulin action. For this reason, it is recommended in the prevention and management of disease states that are characterised by insulin resistance ("metabolic syndrome"). Increased skeletal muscle glucose uptake, as a consequence of carbohydrate ingestion, maintains carbohydrate supply to contracting muscle, at a time when glycogen levels are reduced, and is associated with enhanced performance. Thus, both health and exercise performance are influenced by the metabolism of glucose within skeletal muscle.

Dietary Carbohydrates↗

PubMed1998

Limits to exercise performance in the heat.

Exercise in the heat results in major alterations in circulatory, thermoregulatory and metabolic function. Hyperthermia appears to be the critical determinant of exercise performance in the heat. Thus, strategies that minimise the rise in core temperature during exercise in the heat are likely to contribute to enhanced performance. These include acclimatisation to heat, adequate hydration prior to exercise and ingestion of fluids during exercise, and pre-exercise cooling.

Acclimatization↗

PubMed1998

Glycemic index--a new tool in sport nutrition?

The glycemic index (GI) provides a way to rank foods rich in carbohydrate (CHO) according to the glucose response following their intake. Consumption of low-GI CHO foods may attenuate the insulin-mediated metabolic disturbances associated with CHO intake in the hours prior to exercise, better maintaining CHO availability. However, there is insufficient evidence that athletes who consume a low-GI CHO-rich meal prior to a prolonged event will gain clear performance benefits. The ingestion of CHO during prolonged exercise promotes CHO availability and enhances endurance and performance, and athletes usually chose CHO-rich foods and drinks of moderate to high GI to achieve this goal. Moderate- and high-GI CHO choices appear to enhance glycogen storage after exercise compared with low GI CHO-rich foods. However, the reason for this is not clear. A number of attributes of CHO-rich foods may be of value to the athlete including the nutritional value of the food or practical issues such as palatability, portability, cost gastric comfort, or ease of preparation.

Dietary Carbohydrates↗

PubMed1998

Effect of epinephrine on muscle glycogenolysis during exercise in trained men.

To test the hypothesis that an elevation in circulating epinephrine increases intramuscular glycogen utilization, six endurance-trained men performed two 40-min cycling trials at 71 +/- 2% of peak oxygen uptake in 20-22 degrees C conditions. On the first occasion, subjects were infused with saline throughout exercise (Con). One week later, after determination of plasma epinephrine levels in Con, subjects performed the second trial (Epi) with an epinephrine infusion, which resulted in a twofold higher (P < 0.01) plasma epinephrine concentration in Epi compared with Con. Although oxygen uptake was not different when the two trials were compared, respiratory exchange ratio was higher throughout exercise in Epi compared with Con (0.93 +/- 0.01 vs. 0.89 +/- 0.01; P < 0.05). Muscle glycogen concentration was not different when the trials were compared preexercise, but the postexercise value was lower (P < 0.01) in Epi compared with Con. Thus net muscle glycogen utilization was greater during exercise with epinephrine infusion (224 +/- 37 vs. 303 +/- 30 mmol/kg for Con and Epi, respectively; P < 0.01). In addition, both muscle and plasma lactate and plasma glucose concentrations were higher (P < 0.05) in Epi compared with Con. These data indicate that intramuscular glycogen utilization, glycolysis, and carbohydrate oxidation are augmented by elevated epinephrine during submaximal exercise in trained men.

Adult↗

PubMed1998

Effect of increased blood glucose availability on glucose kinetics during exercise.

This study examined the effect of increased blood glucose availability on glucose kinetics during exercise. Five trained men cycled for 40 min at 77 +/- 1% peak oxygen uptake on two occasions. During the second trial (Glu), glucose was infused at a rate equal to the average hepatic glucose production (HGP) measured during exercise in the control trial (Con). Glucose kinetics were measured by a primed continuous infusion of D-[3-3H]glucose. Plasma glucose increased during exercise in both trials and was significantly higher in Glu. HGP was similar at rest (Con, 11.4 +/- 1.2; Glu, 10.6 +/- 0.6 micromol . kg-1 . min-1). After 40 min of exercise, HGP reached a peak of 40.2 +/- 5.5 micromol . kg-1 . min-1 in Con; however, in Glu, there was complete inhibition of the increase in HGP during exercise that never rose above the preexercise level. The rate of glucose disappearance was greater (P < 0.05) during the last 15 min of exercise in Glu. These results indicate that an increase in glucose availability inhibits the rise in HGP during exercise, suggesting that metabolic feedback signals can override feed-forward activation of HGP during strenuous exercise.

Adult↗

PubMed1998

Muscle metabolites and performance during high-intensity, intermittent exercise.

Six men were studied during four 30-s "all-out" exercise bouts on an air-braked cycle ergometer. The first three exercise bouts were separated by 4 min of passive recovery; after the third bout, subjects rested for 4 min, exercised for 30 min at 30-35% peak O2 consumption, and rested for a further 60 min before completing the fourth exercise bout. Peak power and total work were reduced (P < 0. 05) during bout 3 [765 +/- 60 (SE) W; 15.8 +/- 1.0 kJ] compared with bout 1 (1,168 +/- 55 W, 23.8 +/- 1.2 kJ), but no difference in exercise performance was observed between bouts 1 and 4 (1,094 +/- 64 W, 23.2 +/- 1.4 kJ). Before bout 3, muscle ATP, creatine phosphate (CP), glycogen, pH, and sarcoplasmic reticulum (SR) Ca2+ uptake were reduced, while muscle lactate and inosine 5'-monophosphate were increased. Muscle ATP and glycogen before bout 4 remained lower than values before bout 1 (P < 0.05), but there were no differences in muscle inosine 5'-monophosphate, lactate, pH, and SR Ca2+ uptake. Muscle CP levels before bout 4 had increased above resting levels. Consistent with the decline in muscle ATP were increases in hypoxanthine and inosine before bouts 3 and 4. The decline in exercise performance does not appear to be related to a reduction in muscle glycogen. Instead, it may be caused by reduced CP availability, increased H+ concentration, impairment in SR function, or some other fatigue-inducing agent.

Adenosine Triphosphate↗

PubMed1997

Effect of muscle glycogen availability on maximal exercise performance.

This investigation determined the influence of pre-exercise muscle glycogen availability on performance during high intensity exercise. Nine trained male cyclists were studied during 75 s of all-out exercise on an air-braked cycle ergometer following muscle glycogen-lowering exercise and consumption of diets (energy content approximately 14 MJ) that were either high (HCHO(80% CHO) or low (LCHO-25% CHO) in carbohydrate content. The exercise-diet regimen was successful in producing differences in pre-exercise muscle glycogen contents [HCHO: 578(SEM 55) mmol x kg(-1) dry mass; LCHO: 364 (SEM 58) P < 0.05 mmol x kg(-1) dry mass]. Despite this difference in muscle glycogen availability, there were no between trial differences for peak power [HCHO 1185 (SEM 50)W, LCHO 1179 (SEM 48)W], mean power [HCHO 547 (SEM 5)W, LCHO 554 (SEM 8)W] and maximal accumulated oxygen deficit [HCHO 54.4 (SEM 2.3) ml x kg(-1), LCHO 54.6 (SEM 2.0) ml x kg(-1)]. Postexercise muscle lactate contents (HCHO 95.9 (SEM 4.6) mmol x kg(-1) dry mass, LCHO 82.7 (SEM 12.3) mmol x kg(-1) dry mass, n = 8] were no different between the two trials, nor were venous blood lactate concentrations immediately after and during recovery from exercise. These results would indicate that increased muscle glycogen availability has no direct effect on performance during all-out high intensity exercise.

Adult↗

PubMed1997

Influence of ingested fluid volume on physiological responses during prolonged exercise.

The effect of different rates of fluid ingestion on heart rate, rectal temperature, plasma electrolytes, hormones and performance was examined during prolonged strenuous exercise conducted at 21 degrees C. Seven well-trained males (24 +/- 1 yr; 68.6 +/- 2.9 kg; VO2 peak = 4.69 +/- 0.17 L min-1; mean +/- SEM) cycled for 2 h at 69 +/- 1% VO2 peak while receiving either no fluid replacement (NF), a volume of water estimated to prevent body weight loss (FR-100 = 2.32 +/- 0.10 L 2 h-1) or 50% of this volume (FR-60 = 1.16 +/- 0.05 L 2 h-1). The 2-h exercise bout was followed by a ride to exhaustion at a workload estimated to be 90% VO2 peak. After 2 h of exercise, NF was associated with a 3.2 +/- 0.1% weight loss, while FR-50 and FR-100 resulted in losses of 1.8 +/- 0.1 and 0.1 +/- 0.1%, respectively. Compared with FR-100, heart rate and rectal temperature were elevated (P < 0.05) during the second hour of exercise in NF, with FR-50 intermediate. Reductions in plasma volume during exercise were greater in NF and FR-50, compared with FR-100 and plasma sodium concentration was elevated in NF, decreased slightly in FR-100, with FR-50 intermediate. Plasma renin activity, aldosterone and atrial natriuretic peptide increased to similar extents in the three trials. Plasma vasopressin remained unchanged for FR-100, increased for NF, with intermediate values for FR-50. Exercise time to exhaustion at 90% VO2-peak was longer in FR-100 (328 +/- 93 s) than NF (171 +/- 75 s) with FR-50 (248 +/- 107 s) not significantly different from either FR-100 or NF. In conclusion, the responses of heart rate, rectal temperature, plasma sodium, and vasopressin during, and performance following, prolonged cycling exercise conducted at 21 degrees C are related to the amount of fluid ingested (i.e. the degree of dehydration).

Adult↗

PubMed1997

Identical expression of CD45R isoforms by CD45RC+ 'revertant' memory and CD45RC+ naive CD4 T cells.

Naive and memory CD4 T cells are frequently defined by exon-specific monoclonal antibodies (mAb) which stain (or not) high- or low-molecular-weight (MW) isoforms of the leucocyte common antigen CD45. The link between isoform and the naive/memory designation is complicated by the fact that CD4 T cells with a 'memory' phenotype (CD45RA-, RB-, RC-, or CD45RO+) may revert ('revertants') and re-express the high mw isoform (CD45RA+, RB+, RC+). Isoform expression also changes during normal T-cell development. Furthermore, the picture may be incomplete since an exon-specific mAb will not detect all possible isoforms on a cell. We have used molecular techniques to determine whether revertant CD4 memory T cells were different from naive T cells with respect to CD45R isoform expression. Using the anti-CD45RC mAb OX22 to purify rat lymphocyte subsets, CD45R isoform expression was examined at the mRNA level in CD4 T cells at different stages of development and compared with that of B cells and unseparated lymphocytes. B cells contained abundant message for the highest MW 3-exon isoform ABC, the 2-exon isoforms AB and BC, and the null isoform O. Both immature CD45RC- (i.e. CD4+8- 'single positive' thymocytes, and peripheral Thy-1+ recent thymic emigrants) and mature CD45RC- 'antigen-experienced' CD4 T cells had message for single-exons B, possibly C and for the O exon. In contrast, CD45RC+ CD4 T cells contained mRNA coding for ABC (low level), AB, BC, B, C (low level) and O (low level). Importantly, there was no difference between CD45RC+ T cells that had not seen antigen ('truly native') and CD45RC+ antigen-experienced revertant memory T cells. This observation has implications for understanding long-term immunological memory.

Animals↗

PubMed1997

Progressive increase in glucose transport and GLUT-4 in human sarcolemmal vesicles during moderate exercise.

Muscle glucose uptake increases progressively during moderate-intensity exercise. To elucidate whether this is due to a progressive increase in sarcolemmal glucose transport capacity, nine men exercised for 40 min at 75% maximal oxygen uptake on a bicycle ergometer. Muscle biopsies were obtained from the vastus lateralis at rest (0 min) and after 5 and 40 min of exercise and used for production of sarcolemmal giant (SG) vesicles. SG vesicle glucose transport at 5 mM increased (P < 0.05) by 38 and 93% after 5 and 40 min of exercise, respectively, compared with glucose transport at rest. The SG vesicle GLUT-4 protein content increased (P < 0.05) by 36 and 91% after 5 and 40 min of exercise, respectively, compared with rest. Thus the increase in vesicle glucose transport was accompanied by a similar increase in SG vesicle GLUT-4 protein content. Muscle glucose and glucose 6-phosphate were low at rest, increased (P < 0.05) 2.2- and 2.3-fold, respectively, after 5 min of exercise, and returned to resting values after 40 min of exercise. It is concluded that the progressive increase in muscle glucose uptake during moderate-intensity exercise may be due at least in part to a progressive increase in sarcolemmal glucose transport and GLUT-4 protein content.

Adult↗

PubMed1997

Interactions between muscle glycogen and blood glucose during exercise.

Muscle glycogen and blood glucose are important substrates for contracting skeletal muscle during exercise. The possibility exists for considerable interaction between muscle glycogen and blood glucose and their effects on muscle glucose uptake and glycogenolysis, respectively. Increases in blood glucose availability have little effect on net muscle glycogen utilization during prolonged, strenuous cycling exercise, but this may be dependent upon the mode and intensity of exercise. No data exist on the direct effect of reduced blood glucose levels on muscle glycogen metabolism. In rats, studies using the perfused hindlimb suggest an inverse relationship between muscle glycogen levels and glucose uptake. A similar conclusion can be drawn from a number of human studies albeit, on occasion, from indirect evidence. The influence of muscle glycogen on glucose uptake involves effects on both membrane glucose transport and intracellular glucose metabolism. Such a relationship would, under conditions of adequate muscle glycogen availability, limit muscle glucose uptake, thereby preserving the relatively small blood glucose supply for the brain, nerves, and blood cells, which are dependent on it for their energy metabolism.

Animals↗

PubMed1996

A method for measuring sarcoplasmic reticulum calcium uptake in the skeletal muscle using Fura-2.

We have presented an assay for measuring the rate of sarcoplasmic reticulum (SR) Ca2+ uptake and Ca2+ release in skeletal muscle homogenates using the fluorescent Ca2+ probe Fura-2. Using this assay, we investigated the effects of an elevated temperature (40 degrees C) and lowered pH (6.8), two factors proposed to be involved in skeletal muscle fatigue, on SR Ca2+ uptake. The EDL muscle was found to have a higher rate of Ca2+ uptake than the soleus (34%). Exposure of the muscles to a raised temperature, but not a reduced pH, resulted in a reduction in the rate of Ca2+ uptake in both the EDL and soleus homogenates. This uptake process was blocked by cyclopiazonic acid (CPA) a specific inhibitor of the major transport protein of the sarcoplasmic reticulum, the Ca(2+)-ATPase. Calcium release was induced using AgNO3 after loading of the vesicles during the uptake process. It was found that AgNO3 was only effective in producing Ca2+ release in the EDL muscles. The soleus muscles did not release Ca2+ under varying [Mg2+] or with Hg2+ substitution for Ag+, suggesting that fast- and slow-twitch muscle fibres require different conditions for maximum Ca2+ release, or that different isoforms of the Ca2+ release channels are present in the different fibres.

Animals↗