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Selected AbstractsNo relationship between enzyme activity and structure of nucleotide binding site in sarcoplasmic reticulum Ca2+ -ATPase from short-term stimulated rat muscleACTA PHYSIOLOGICA, Issue 4 2009T. Mishima Abstract Aim:, We examined whether structural alterations to the adenine nucleotide binding site (ANBS) within sarcoplasmic (endo) reticulum Ca2+ -ATPase (SERCA) would account for contraction-induced changes in the catalytic activity of the enzyme as assessed in vitro. Methods:, Repetitive contractions were induced in rat gastrocnemius by electrical nerve stimulation. Measurements of sarcoplasmic reticulum properties were performed on control and stimulated muscles immediately after or at 30 min after the cessation of 5-min stimulation. In order to examine the properties at the ANBS, the binding capacity of SERCA to fluorescence isothiocyanate (FITC), a competitive inhibitor at the ANBS, was analysed in microsomes. Results:, Short-term electrical stimulation evoked a 23.9% and 32.6% decrease (P < 0.05) in SERCA activity and in the FITC binding capacity, respectively, in the superficial region of the muscle. Whereas SERCA activity reverted to normal levels during 30-min recovery, a restoration of the FITC binding capacity did not occur. Conclusion:, The discordant changes between the enzyme activity and the FITC binding suggest that, at least during recovery after exercise, changes in SERCA activity may not correlate closely with structural alterations to the ANBS within the enzyme. [source] Muscle, tendon, and somatotropin responses to the restriction of muscle blood flow induced by KAATSU-walk trainingEQUINE VETERINARY JOURNAL, Issue S36 2006T. ABE Summary Objective: The efficacy of KAATSU training has been demonstrated in human athletes, both as a therapeutic method as well as a training aid. The purpose of this study was to investigate the effects of slow walk training combined with restriction of muscle blood flow (KAATSU) on muscle and tendon size. Methods: Six healthy, unfit Standardbred mares performed walking (240 m/min for 10 min and then 5 min recovery) with KAATSU, and 6 mares performed walking without KAATSU. A specially designed elastic cuff1 was placed at the most proximal position of the forelegs and inflated to a pressure of 200,230 mmHg throughout the walking and recovery sessions. The training was conducted once a day, 6 days/week for 2 weeks. Skeletal muscle thickness and tendon thickness were measured using B-mode ultrasound at baseline and after 2 weeks of training. Venous blood samples were obtained before the first acute exercise and 5, 15 and 60 min afterwards. Serum somatotropin concentration was determined using a commercially available equine-specific ELISA kit. Results: The acute increase in plasma somatotropin was 40% greater (P<0.05) in the KAATSU-walk group than in the Control-walk group 5 min after exercise and remained elevated (P<0.05) at 15 and 60 min post exercise compared with the Control-walk group. After 2 weeks of training, muscle thickness increased (P<0.05) 3.5% in the KAATSU-walk group but did not change in the Control-walk group (0.7%). Tendon thickness did not change (P>0.05) in either group. Conclusions: These data demonstrate that KAATSU training can induce muscle hypertrophy in horses and suggest that KAATSU training may provide significant therapeutic/rehabilitative value in horses, as has been shown in man. [source] Application of a constant blood withdrawal method in equine exercise physiology studiesEQUINE VETERINARY JOURNAL, Issue 6 2001P. BARAGLI Summary The aim of the present study was to test a constant blood withdrawal method (CBWM) to collect blood samples from horses during treadmill exercise. CBWM was performed in 4 Standardbreds and 5 Haflinger horses. A peristaltic pump was used to control blood aspiration from an i.v. catheter via an extension line. Blood was collected using an automatic fractions collector, with a constant delay time between the drawing of blood and sample collection. Blood withdrawal using CBWM was made during a treadmill standardised exercise test (SET). A blood flow of 12 ml/min was used and samples collected every 60 s during the entire period of exercise. The volume of blood collected in each sample tube was 12.1 ± 0.2 ml, with a delay time of mean ± s.d. 25.3 ± 0.8 s. Plasma lactate kinetics based on measurement of lactate in each fraction showed an exponential increase during the first 13 min of exercise (10.5 min of SET and 2.5 min recovery). The peak plasma lactate concentration was observed between 2.5 and 5.5 min after the end of SET. CBWM permits the kinetics of lactate and other blood-borne variables to be studied over time. This method could be a valuable aid for use in studying equine exercise physiology. [source] Selective Long-Term Electrical Stimulation of Fast Glycolytic Fibres Increases Capillary Supply but not Oxidative Enzyme Activity in Rat Skeletal MusclesEXPERIMENTAL PHYSIOLOGY, Issue 5 2000S. Egginton Glycolytic fibres in rat extensor digitorum longus (EDL) and tibialis anterior (TA) were selectively activated, as demonstrated by glycogen depletion, by indirect electrical stimulation via electrodes implanted in the vicinity of the peroneal nerve using high frequency (40 Hz) trains (250 ms at 1 Hz) and low voltage (threshold of palpable contractions). This regime was applied 10 times per day, each bout being of 15 min duration with 60 min recovery, for 2 weeks. Cryostat sections of muscles were stained for alkaline phosphatase to depict capillaries, succinate dehydrogenase (SDH) to demonstrate oxidative fibres, and periodic acid-Schiff reagent (PAS) to verify glycogen depletion. Specific activity of hexokinase (HK), 6-phosphofructokinase, pyruvate kinase, glycogen phosphorylase and cytochrome c oxidase (COX) were estimated separately in homogenates of the EDL and the predominantly glycolytic cortex and oxidative core of the TA. Stimulation increased the activity of HK but not that of oxidative enzymes in fast muscles. Comparison of changes in oxidative capacity and capillary supply showed a dissociation in the predominantly glycolytic TA cortex. Here, COX was 3.9 ± 0.68 ,M min-1 (g wet wt)-1 in stimulated muscles compared with 3.7 ± 0.52 ,M min-1 (g wet wt)-1 in contralateral muscles (difference not significant), while the percentage of oxidative fibres (those positively stained for SDH) was also similar in stimulated (14.0 ± 2.8%) and contralateral (12.2 ± 1.9%) muscles. In contrast, the capillary to fibre ratio was significantly increased (2.01 ± 0.12 vs. 1.55 ± 0.04, P < 0.01). We conclude that capillary supply can be increased independently of oxidative capacity, possibly due to haemodynamic factors, and serves metabolite removal to a greater extent than substrate delivery. [source] The temperature response of photosynthesis in tobacco with reduced amounts of RubiscoPLANT CELL & ENVIRONMENT, Issue 4 2008DAVID S. KUBIEN ABSTRACT The reasons for the decline in net CO2 assimilation (A) above its thermal optimum are controversial. We tested the hypothesis that increasing the ratio of Rubisco activase to Rubisco catalytic site concentration would increase the activation state of Rubisco at high temperatures. We measured photosynthetic gas exchange, in vivo electron transport (J) and the activation state of Rubisco between 15 and 45 °C, at 38 and 76 Pa ambient CO2, in wild-type (WT) and anti- rbcS tobacco. The Rubisco content of the anti- rbcS lines was 30% (S7-1) or 6% (S7-2) of WT, but activase levels were the same in the three genotypes. Anti- rbcS plants had lower A than WT at all temperatures, but had a similar thermal optimum for photosynthesis as WT at both CO2 levels. In WT plants, Rubisco was fully activated at 32 °C, but the activation state declined to 64% at 42 °C. By contrast, the activation state of Rubisco was above 90% in the S7-1 line, between 15 and 42 °C. Both A and J declined about 20% from Topt to the highest measurement temperatures in WT and the S7-1 line, but this was fully reversed after a 20 min recovery at 35 °C. At 76 Pa CO2, predicted rates of RuBP regeneration-limited photosynthesis corresponded with measured A in WT tobacco at all temperatures, and in S7-1 tobacco above 40 °C. Our observations are consistent with the hypothesis that the high temperature decline in A in the WT is because of an RuBP regeneration limitation, rather than the capacity of Rubisco activase to maintain high Rubisco activation state. [source] Improving exercise capacity, 6 wk training tends to reduce circulating endothelin after heart transplantationCLINICAL TRANSPLANTATION, Issue 6 2004Stèphane Doutreleau Abstract:, Short-term survival is no longer the pivotal issue after heart transplantation but, most heart-transplant (Htx) patients still present with increased circulating endothelin-1 (ET) and reduced exercise capacity. ET-1 limits both exercise-induced vasodilation and blood flow redistribution toward acting muscles and might be accessible to training. This study was performed to investigate the effect of training on ET-1 and whether an eventual training-induced improvement in exercise capacity may be related to reduced baseline or exercise circulating ET-1 in Htx patients. Five Htx patients performed a maximal bicycle exercise test and an endurance exercise test before and after a training program of 18 exercises sessions during 6 wk. ET-1 was determined by radioimmunoassay at rest, end endurance exercise and 30 min recovery, before and after training. Training improved significantly Htx's maximal oxygen uptake (+13.1 ± 4.8%; p < 0.05) and also reduced significantly the endurance exercise-induced heart rate increase. Resting ET-1 was increased in Htx (5.98 ± 1.88 vs. 1.61 ± 0.25 pmol/L in controls; p < 0.01) but although ET-1 modulation might participate in training-induced beneficial effects, training failed to modulate either resting or exercise ET-1 plasma level. Training-induced improvement in exercise capacity might not mainly due to decreased ET-1 after heart transplantation. Further supporting the usefulness of training, these preliminary data suggest that improved exercise capacity may not be mainly due to decreased ET-1 in Htx patients. Further, larger scale studies will be needed to investigate whether an impaired nitric oxide pathway stimulation might explain such results and whether a longer training program can reduce local ET-1, arising from working muscles after heart transplantation. [source] |