Biomedical subjects

A N Belcastro

Publications and source records attributed to A N Belcastro.

At least 19 recordsLinked to original sources

PubMed1992

Intraluminal Ca2+ dependence of Ca2+ and ryanodine-mediated regulation of skeletal muscle sarcoplasmic reticulum Ca2+ release.

The action of ryanodine upon sarcoplasmic reticulum (SR) Ca2+ handling is controversial with evidence for both activation and inhibition of SR Ca2+ release. In this study, the role of the intraluminal SR Ca2+ load was probed as a potential regulator of ryanodine-mediated effects upon SR Ca2+ release. Through dual-wavelength spectroscopy of Ca2+:antipyrylazo III difference absorbance, the intraluminal Ca2+ dependence of ryanodine and Ca(2+)-induced Ca2+ release (CICR) from skeletal SR vesicles was examined. Ryanodine addition after initiation of Ca2+ uptake (a) increased the intraluminal Ca2+ sensitivity of CICR and (b) stimulated spontaneous Ca2+ release with a delayed onset. These ryanodine effects were inversely proportional to the intraluminal Ca2+ load. Ryanodine also inhibited subsequent CICR after reaccumulation of Ca2+ released from the initial CICR. These results provide evidence that ryanodine inhibits transitions between low and high affinity Ca2+ binding states of an intraluminal Ca2+ compartment, possibly calsequestrin. Conformational transitions of calsequestrin may be reciprocally coupled to transitions between open and closed states of the Ca2+ release channel.

Adenosine Triphosphate↗

PubMed1992

Calcium-activated neutral protease effects upon skeletal muscle sarcoplasmic reticulum protein structure and calcium release.

In this study, the effects of Ca(2+)-activated neutral protease (CANP) upon skeletal muscle heavy sarcoplasmic reticulum (HSR) structure and function were investigated. CANP was immunolocalized to the 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid detergent-insoluble fraction of purified HSR membranes. Ca2+ activation of the endogenous membrane-bound CANP produced a characteristic partial fragmentation of the HSR 565-kDa Ca2+ release channel. Similarly, the major substrate for both micromolar and millimolar Ca(2+)-sensitive isoforms of exogenous CANP was the Ca2+ release channel with proteolysis of a 88-kDa HSR protein also observed. Ca2+ release channel proteolysis was initiated at a single cleavage site with coincidental production of 410- and 150-kDa peptide fragments. Appearance of 160- and 137-kDa limiting peptides accompanied secondary proteolysis of the primary 410- and 150-kDa fragments, respectively. Despite extensive proteolysis of the Ca2+ release channel, CANP did not dramatically alter the Ca2+ handling and ryanodine binding properties of HSR membranes. The association of CANP with isolated HSR membranes suggests that, in vivo, this protease may modify an additional property of the Ca2+ release channel. This may be related to the CANP-susceptible structural association of the Ca2+ release channel with dihydropyridine receptors at T-tubule/sarcoplasmic reticulum junctions.

Adenosine Triphosphate↗

PubMed1992

Effect of manipulation of plasma lactate on integrated EMG during cycling.

This investigation was undertaken to record electromyographic activity of the vastus lateralis muscle during incremental cycling exercise and to determine whether it would be sensitive to altered dynamics of plasma lactate increases seen with intense exercise. Trained cyclists (N = 6) performed two progressive, stepwise exercise tests (23.5 W.min-1) to fatigue on a cycle ergometer at 90 rpm. One of the exercise tests was preceded by arm ergometer exercise in an attempt to elevate the circulating plasma lactate levels prior to starting the criterion exercise test. The starting mean plasma lactate values were 4.59 and 26.69 mmol lactate.-1 for the two exercise sessions. Cardiorespiratory values did not differ significantly between exercise sessions completed in the absence and presence of increased circulating plasma lactate. The no-arm trial (i.e., nonelevated plasma lactate condition) was associated with a plasma lactate inflection point (Tlac) at 72.6% VO2max. Previous arm exercise elevated the lactate such that during the criterion exercise plasma lactate values were decreasing with increasing power output at lower exercise intensities. As exercise intensity increased lactate values also increased beginning at a power output of about 76% VO2 max. Mean per cycle integrated EMG (CIEMG) increased linearly with increased power output in both exercise sessions. The slopes of the EMG-power output curve were not significantly different (P less than 0.05). There were no inflection points in these curves. The absence of an inflection point show that surface EMG does not provide an indication of Tlac.

Adolescent↗

PubMed1991

Regulation of ATP-stimulated releasable myofilaments from cardiac and skeletal muscle myofibrils.

The mechanism underlying the formation of easily releasable myofilaments, from myofibrils treated with an ATP-containing relaxing solution, was examined in this investigation. The proportion of releasable myofilaments purified from myofibrils of cardiac, fast- and slow-twitch muscles increased as the [ATP] was raised from 0 to 8.5 mM. The protein composition of the easily releasable myofilaments did not differ with increasing ATP concentrations as observed by 5-15% linear gradient SDS-PAGE. There is a nucleotide specificity to the release of myofilaments in the order of ATP greater than GTP much greater than UTP greater than CTP. Experiments with AMP-PNP and inorganic phosphate (Pi) showed that ATP hydrolysis and the build up of Pi are not requirements in the formation of the easily releasable myofilaments. The release of myofilaments was found to be insensitive to variations in pH from 6.5 to 7.5. The ATP stimulation of myofilaments release is ubiquitin-independent, since incubation of purified myofibrils with ubiquitin (1-100 micrograms/ml) at both 20 and 37 degrees C did not change the amount released. Modifying the free sulfhydryl group content by treatment of myofibrils with NEM (0.01-1 mM) or silver nitrate (0.1-10 mM) decreased the proportion of myofilaments that were releasable. Exclusion of 1 mM DTT from the preparation of myofibrils had similar results. These results indicate that the formation of easily releasable myofilaments can be mediated by metabolically related parameters such as the adenosine nucleotides and the reduction-oxidation status of the myofibrillar proteins of striated muscle.

Actin Cytoskeleton↗

PubMed1991

Biochemical adaptation of cardiac and skeletal muscle to physical activity.

1. Female Wistar rats were randomly assigned to control (C) or exercising (T) groups and subsequently portioned into 1, 3, 5 and 10 day T and C groups. The T groups completed a progressive endurance running program. Biochemical indices of adaptation were measured in cardiac muscle and in plantaris and soleus muscles of C and T animals after their last exercise bout. 2. In cardiac muscle, myofibrillar ATPase activity was significantly elevated in the 3T (0.241 +/- 0.031) and 5T (0.242 +/- 0.013) groups (P less than or equal to 0.05) compared to their respective controls (3C = 0.187 +/- 0.015 and 5C = 0.190 +/- 0.007). 3. After 10 days of training cardiac myofibrillar ATPase activity was elevated by 17% but this was not significant (P greater than or equal to 0.05). 4. No changes in myofibrillar ATPase activity were seen in skeletal muscle (P greater than or equal to 0.05), however, hexokinase activity progressively increased and was significantly elevated in the 3T, 5T and 10T soleus and plantaris muscles of rats over controls (P less than or equal to 0.05). 5. Minimal nonsignificant changes were noted in the hexokinase activity of the hearts of all T groups (P greater than or equal to 0.05). 6. These results indicate that metabolic adaptation of the heart and skeletal muscles takes place after as little as three training sessions. 7. Although the adaptation of the skeletal muscles continually progresses, the adaptation of the heart appears to be transitory.(ABSTRACT TRUNCATED AT 250 WORDS)

Adaptation, Physiological↗

PubMed1990

Improved resolution of the initial fast phase of heavy sarcoplasmic reticulum Ca2+ uptake by Ca2+:antipyrylazo III dual-wavelength spectroscopy.

The effect of ATP upon difference absorbance due to Ca2+ and Mg2+ complexation with the metallochromic dye, Antipyrylazo III (AP III), was investigated. At divalent cation concentrations appropriate for Sarcoplasmic Reticulum Ca2+ transport, wavelengths (greater than 670 nm) were found whereupon the addition of up to 1mM nucleotide did not alter divalent cation:AP III difference absorbance. At these sample wavelengths an initial rapid uptake of Ca2+ by Heavy SR (HSR) was clearly resolved by dual wavelength spectroscopy of Ca2+:dye difference absorbance. Elimination of ATP interference of Ca2+:AP III absorbance by Mg2+ elevation (3-10mM) was shown to be an inappropriate general strategy for AP III spectroscopic studies of HSR Ca2+ transport due to Mg2+ inhibition of ryanodine receptor mediated Ca2+ release.

Adenosine Triphosphate↗

PubMed1990

Glucoregulatory response to moderate exercise in long-term islet cell autografted dogs.

The glucoregulatory response to moderate treadmill exercise (approximately 60% maximum heart rate; 60 min at 100 m/min, 12% grade) was examined in six controls and six pancreatectomized dogs that had been normoglycemic and insulin independent for more than 1 year since autograft of isolated islets of Langerhans (Tx). There were no significant intergroup differences in plasma glucose levels during exercise, but return to baseline after exercise was delayed in Tx (p less than 0.05). In Tx, the initially suppressed insulin levels rose above baseline from 30 to 60 min. Within Tx, exercise-induced levels of plasma glucagon and epinephrine were more variable than control and strongly correlated (r = 0.81, p less than 0.001), perhaps indicating that the A cells within the grafted islets were regulated by circulating beta-adrenergic agonists. We conclude that the isolated islets were removed from direct central control. In Tx dogs, the variable counterregulatory responses and the diminished recovery of plasma glucose after exercise indicate reliance on alternative glucoregulatory mechanisms.

Adrenal Glands↗

PubMed1989

Time dependent response of cardiac myofibrillar ATPase activity to exercise.

1. The purpose of the present study was to investigate the time course of run training effects on the Ca2+ kinetics of the cardiac myofibrillar ATPase activity in female Sprague-Dawley rats. 2. The cardiac myofibrillar ATPase activity was measured at varying Ca2+ levels, and the Hill-n and pCa50 were measured in the hearts of rats after 3, 6 and 9 weeks of running training with a training program that began with an initially high intensity (HINT) and a training program with a more progressive increase in intensity (PROG). 3. After 3 and 6 weeks of training cardiac myofibrillar ATPase activity in the hearts of the trained rats in both training programs was elevated by 28-40% over the control group (P less than 0.05) at a pCa5 but was not different from the control groups after 9 weeks of training (P greater than or equal to 0.05). 4. Also the Ca2+ co-operativity as measured by the Hill-n was elevated in the hearts of the trained rats after 6 and 9 weeks of training when compared to control groups suggesting changes in the regulatory proteins of the myofibrils of hearts from trained rats. 5. The elevations in cardiac myofibrillar ATPase activity suggest that the myocardium responded to the training stimulus in a phasic manner. 6. The regression of cardiac myofibrillar ATPase in the late weeks of training might be related to a reduction or a loss of a specific training stimulus for the myocardium.

Adenosine Triphosphatases↗

PubMed1989

Dietary carbohydrate-to-fat ratio: influence on whole-body nitrogen retention, substrate utilization, and hormone response in healthy male subjects.

The effect of a high-carbohydrate and a high-fat diet on nitrogen retention, substrate utilization, and serum hormone concentrations was assessed in six healthy male subjects. Both diets were fed at a level estimated to provide maintenance and 75% maintenance energy requirements. Urine and feces were collected and analyzed for N and energy content. Anthropometric measurements; fasting and postprandial oxygen consumption; and serum levels of glucose, triglycerides, and metabolic hormones were measured. The high-fat diet increased N retention at both energy levels with significance reached at maintenance energy intakes (p less than 0.05). The high-fat diet resulted in less weight loss (p less than 0.05) at low energy and a consistently lower respiratory quotient (p less than 0.05), indicative of increased fat oxidation. The N sparing effect of the high-fat diet did not appear to be explained by hormone levels observed but may be substrate mediated.

Adult↗

PubMed1989

Effects of strenuous maternal exercise on fetal organ weights and skeletal muscle development in rats.

The purpose of the present study was to observe the effects of strenuous maternal aerobic exercise throughout gestation on fetal outcome in the rat. The strenuous exercise intensity consisted of a treadmill speed of 30 m.min-1 on a 10 degrees incline, for 120 min.day-1, 5 days.week-1. The rats were conditioned to run on a motor-driven treadmill by following a progressive two-week exercise program, so that by the end of the two weeks the rats were capable of running comfortably at this strenuous intensity in the non-pregnant state. Following the two-week running programme, the rats were paired by weight and randomly assigned to either a pregnant group that continued the running program throughout gestation (pregnant runner), or a pregnant group that did not continue the running program throughout pregnancy (pregnant control). At birth the neonates born to the pregnant running group did not differ in average neonatal body weight values, number per litter or total litter weight values when compared to controls, nor were superficial gross abnormalities observed in neonates born to the pregnant control or pregnant running groups. The strenuous maternal exercise intensity did not alter neonatal organ weight values (brain, heart, liver, lung, kidney), nor neonatal skeletal muscle (gastrocnemius, sternomastoid, diaphragm) when compared to control values. It is suggested that maternal exercise of this intensity throughout gestation does not affect fetal outcome in the rat, and may be due to the animals accustomization to the strenuous exercise protocol prior to pregnancy.

Animals↗

PubMed1989

The influence of high-velocity circuit resistance training on VO2max and cardiac output.

In order to investigate the influence of high-velocity circuit resistance training on maximal aerobic power, maximal stroke volume and cardiac output, and blood lactate removal during recovery, 16 habitually active males were blocked on initial VO2max into either training or control groups. The training group completed two (weeks 1 and 2) or three (weeks 3-6) circuits of 10 variable-resistance hydraulic exercise stations at an exercise: relief ratio of 1:2 on alternate days over six weeks. Angular velocities of movement were maintained at approximately 3.1 rad.s-1. Following training, the VO2max was increased (p less than .01) from 4.32 to 4.68 1.min-1. Maximal stroke volume was increased (p less than .05) from 120 to 129 mL and heart rate response to an absolute submaximal exercise load was decreased (p less than .05) from 153 to 146 beats.min-1. As well, enhanced (p less than .01) removal of lactate from the blood was observed during recovery from exhausting exercise. No changes were observed for control subjects. These results indicate that positive alterations in aerobic and cardiovascular function may be achieved consequent to high-velocity circuit resistance training.

Cardiac Output↗

PubMed1988

Influence of exercise on cardiac and skeletal muscle myofibrillar proteins.

The purpose of this study was to examine the Ca2+-Mg2+ myofibrillar ATPase and protein composition of cardiac and skeletal muscle following strenuous activity to voluntary exhaustion. Sprague-Dawley rats (200 g) were assigned to a control and exercised group, with the run group completing 25 m.min-1 and 8% grade for 1 hour. Following activity, the myocardial Ca2+-Mg2+ myofibrillar ATPase activity -pCa relationship had undergone a rightward shift in the curve. Electrophoretic analysis revealed a change in the pattern of cardiac myofibrillar protein bands, particularly in the 38-42 Kdalton region. Enzymatic analysis of myofibrillar proteins from plantaris muscle, revealed no change in Ca2+ regulation following exercise. Electronmicrographic and electrophoretic analysis revealed extensively disrupted sarcomeric structure and a change in the ratio of several plantaris myofibrillar proteins. No difference was observed for myosin: Actin: tropomyosin ratios; however a dramatic reduction in 58 and 95 Kdalton proteins were evident. The results indicate that prolonged running is associated with similar responses in cardiac and skeletal muscle myofibrillar protein compositions. The abnormalities in myofibrillar ultrastructure may implicate force transmission failure as a factor in exercised-induced muscle damage and/or fatigue.

Animals↗

PubMed1988

An improved method for the isolation of rat cardiac sarcoplasmic reticulum.

Preparations of cardiac sarcoplasmic reticulum (CSR) isolated from the rat by differential centrifugation have been widely used for measuring alterations in intracellular calcium flux in response to metabolic and pharmacologic disruptions. However, the purity of these SR fractions has not been firmly established. Using a combination of differential and linear sucrose gradient centrifugation, we have isolated rat CSR with high specific activity and purity. By SDS-PAGE analysis, the preparation is enriched in a protein (110 kD) of similar size to the Ca2+-ATPase of SR from other sources. Gels stained with the dye 'Stains-All' reveal a blue colored 55 kD band, confirming the presence of calsequestrin, the intraluminal low-affinity calcium binding protein of SR. The presence of the transmembrane 53 kD glycoprotein of SR was confirmed by endoglycosidase-H treatment followed by SDS-PAGE and also by a modified Western blotting technique. The rate of calcium uptake in this preparation averages 130 nmol/mg over the first minute of accumulation, approximately 4 times that previously reported for rat CSR. Calcium uptake in our preparation was essentially complete within 5 minutes. Preparations isolated by this method should be of value in future studies measuring alterations in rat CSR function.

Acetylglucosaminidase↗

PubMed1987

The role of intermediary metabolism in the maintenance of proton and charge balance during exercise.

The purpose of this investigation was to examine the role of intermediary metabolism in the maintenance of proton and charge balance in rainbow trout white muscle during exercise. With increasing power outputs, there was a greater reliance on white fibers and anaerobic processes for energy production. Glycogen content declined from a pre-exercise (pre-ex) level of 23 to less than 1 mumol/g following the exhaustive swim, with its greatest rate of decline occurring during the burst swim. Lactate accumulation reached a maximum of 43 mumol/g during the exhaustive swim. PCr declined from about 20 to less than 2 mumol/g at exhaustion with a concomitant accumulation of Cr. ATP decreased from about 7.3 to 2.7 mumol/g while inorganic phosphate and IMP increased to about 56 and 4.3 mumol/g, respectively. The intramuscular pH fell from 6.97 to 6.93 during the sustained swim, declining further to 6.65 during the burst swim and reaching a minimum of 6.56 at exhaustion. Exercise induced depletions of high energy compounds and accumulations of metabolic end products nearly stabilized the accompanying intracellular perturbations in charge and proton levels. Compensatory shifts in Na+, K+ and Cl- served to negate the residual imbalances such that electrical neutrality, membrane potential and pH were preserved.

Adenine Nucleotides↗

PubMed1987

Are indices of free radical damage related to exercise intensity.

The possibility that plasma levels of malonaldehyde (MDA) are altered by exercise has been examined. The presence of MDA has been recognized to reflect peroxidation of lipids resulting from reactions with free radicals. Maximal exercise, eliciting 100% of maximal oxygen consumption (VO2max) resulted in a 26% increase in plasma MDA (P less than 0.005). Short periods of intermittent exercise, the intensity of which was varied, indicated a correlation between lactate and MDA (r2 = 0.51) (p less than 0.001). Blood lactate concentrations increased throughout this exercise regimen. A significant decrease (10.3%) in plasma MDA occurred at 40% VO2max. At 70% VO2max plasma MDA was still below resting values, however the trend to an increase in MDA with exercise intensity was evident. At exhaustion, plasma MDA and lactate were significantly greater than at rest. These results suggest, that exhaustive maximal exercise induces free radical generation while short periods of submaximal exercise (i.e. less than 70% VO2max) may inhibit it and lipid peroxidation.

Adult↗

PubMed1987

Myofibril and sarcoplasmic reticulum changes during muscle development: activity vs inactivity.

1. The purpose of this study was to determine whether biochemical changes of skeletal muscle that occur as a result of exercise in young rats persist into adulthood. 2. Littermates (10 days old) were assigned to a 3, 6 and 12 week control or training group. In addition, a rest-exercise group (R-E) and exercise-rest (E-R) group were included. 3. The rest-exercise and exercise-rest rats were maintained for the 12 weeks with the first 6 weeks being either rest or exercise and the condition reversed during the last 6 weeks of the experiment. 4. Myofibril ATPase activity of rat plantaris increased from the 10d to 12 week animals (P less than 0.05). As anticipated, training resulted in a lowered activity at 6 and 12 weeks compared to controls. 5. The Ca2+ uptake and Ca2+-ATPase activity of the sarcoplasmic reticulum followed a similar pattern. 6. With regard to the exercise-rest rats, the myofibril and SR ATPase activities at 12 weeks were comparable to the 12 weeks control rats. 7. The rest-exercise group approximated the 12 week training group with regard to myofibril and SR ATPase activities (P less than 0.05). 8. The results suggest that the training adaptations that occur during development of skeletal muscle return to normal, when training ceases in the adult rat. 9. Furthermore, animals that started to train prior to puberty do not have a greater capacity to adapt than animals which initiated training during adulthood.

Aging↗

PubMed1987

Decrease in serum testosterone levels with maximal intensity swimming exercise in trained male and female swimmers.

Circulating testosterone levels have generally been reported to increase during short term, intense running or bicycle ergometer exercise. Swimming differs from such activity because of the strenuous arm activity and because it is conducted in a liquid medium in the horizontal position with the weight totally supported. To investigate whether such differences modify the serum testosterone response to exercise, we measured serum testosterone levels before and after a maximal intensity swimming test in ten elite male and ten elite female swimmers. Levels of circulating testosterone fell in 19 of 20 swimmers in contrast to previous reports of exercise in the vertical position. In women mean testosterone levels declined by 39.4% from baseline values and in males, mean testosterone levels declined by 19.0%. Since the incremental protocol was similar to in design to maximal intensity tests conducted on a treadmill or bicycle ergometer, these data suggest that the differing physical circumstances of swimming lead to a qualitatively different testosterone response to exercise.

Adolescent↗

PubMed1987

Insulin and islet cell transplants: effects on diabetic rat cardiac myofibril ATPase.

Streptozotocin-induced diabetes results in depression of growth rate, cardiac myofibril ATPase activity, and elevated plasma glucose levels. Reversibility of these changes with daily insulin injections and pancreatic islet cell transplants was investigated and compared. Cardiac myofibril ATPase activities (mumol Pi X mg-1 X min-1) were depressed in the uncontrolled diabetic (D) group over the complete range of Ca2+ concentrations tested (e.g., 0.057 +/- 0.017 at 10 microM free Ca2+) with respect to the control (C) group (0.113 +/- 0.009). Neither the transplanted (T) group (0.128 +/- 0.017) nor the insulin injected (I) group (0.111 +/- 0.014) was significantly different from the C animals. Normal growth rates were restored in both I and T groups, whereas in the D group weight gains were negligible in comparison. Cardiac myofibril protein yields (in mg/g wet wt) were not significantly different among groups. These findings indicate that both islet cell transplants and daily insulin injections are capable of normalizing plasma glucose levels, cardiac myofibril ATPase activity, and growth rates in STZ-diabetic rats.

Adenosine Triphosphatases↗