Membrane conductance in trained and untrained subjects using either steady state or single breath measurements of NO transfer.

The aim of this work was to define the relationship between membrane conductance for NO (Dm) and physical activity by using either the steady state NO transfer (T(LNO)SS) or the single breath method (T(LNO)SB), making the hypothesis that NO transfer is only limited by the membrane. Alterations in T(LNO)SS with lung volume during tidal ventilation were measured in six subjects at rest and during steady exercise at 30, 60, and 80% of maximal aerobic power (MAP). A fast responding chemoluminescent NO analyser was used. Two calculation methods were used by sampling NO: (1) at mid-tidal volume, (2) in the middle of the alveolar plateau. T(LNO)SB at rest and maximal oxygen consumption (V(.-)O(2)max) were also measured in 18 other subjects. At rest T(LNO)SS with method 2 was 192% of the value given by method 1. T(LNO)SS with method 1 increased by 50% with 80% MAP as it did not change with method 2. Method 2 seemed inaccurate. T(LNO)SB at rest, which is closely related to Dm, was correlated to age and V(.-)O(2)max, T(LNO)SB=182-1.2 age+24.3 V(.-)O(2) max(l min(-1)) (p<0.01, r(2)=0.72). The T(LNO)SS and T(LNO)SB versus lung volume relationships suggest an influence of the breathing pattern on Dm. Dm can be estimated either by these two NO transfer methods, however the use of the T(LNO)SS method is highly sensitive to the alveolar sampling level. Dm increase during exercise is a function of MAP. Dm at rest decreases with age as it increases with MAP.     

Method for diagnosis and control of aerobic training in rats based on lactate threshold

We propose a protocol for determination of lactate threshold (LT) and test the validity of one aerobic training based on LT in rats. In group I, V(LTi) (velocity at LT before training) was determined in all rats (n=10), each rat training at its own V(LTi) and in group II, animals (n=7) ran at 15 m min(-1), the mean V(LTi) of group I. The training consisted of daily runs at V(LTi) for 50 min, 5 days/week, for 4 weeks. In group I, this program increased V(LT) (V(LTi) 14.90+/-1.49 m min(-1) and V(LTf), after training, 22.60+/-1.17 m min(-1)) and the velocity at exhaustion (19.50+/-1.63 m min(-1) and 27.60+/-1.17 m min(-1)). [Lactate] at LT (2.62+/-0.43 mmol L(-1) versus 2.11+/-0.15 mmol L(-1)) and relative values of LT (76+/-3% versus 82+/-2%) stayed unaltered. In group II the V(LTf) was 20+/-1.8 m.mim(-1), the [lactate] at the LT, 2.02+/-0.17 mmol.L(-1); the exhaustion speed, 23.57+/-2.11 m.mim(-1) and relative value of LT, 82.71+/-2.29%. There were no significant differences in these parameters between groups I and II. Thus, this protocol based on LT is effective and the mean V(LT) determined in a small number of healthy untrained rats can be used for aerobic training in a larger group of healthy animals of same gender and age.     

The effect of regular physical activity on the left ventricle systolic function in patients with chronic coronary artery disease

The purpose of this study was to assess the influence of aerobic training on the left ventricular (LV) systolic function. Thirty patients with stable coronary artery disease, who had participated in the conducted 3-month physical training, were retrospectively divided into 2 cohorts. While patients in the cohort I (n=14) had continued training individually for 12 months, patients in the cohort II (n=16) had stopped training after finishing the conducted program. Rest and stress dobutamine/atropine echocardiography was performed in all patients before the training program and 1 year later. The peak systolic velocities of mitral annulus (Sa) were assessed by tissue Doppler imaging for individual LV walls. In addition, to determine global LV systolic longitudinal function, the four-site mean systolic velocity was calculated (Sa glob). According to the blood supply, left ventricular walls were divided into 5 groups: A- walls supplied by nonstenotic artery; B- walls supplied by coronary artery with stenosis ≤50 %; C- walls supplied by coronary artery with stenosis 51-70 %; D- walls with stenosis of supplying artery 71-99 %; and E- walls with totally occluded supplying artery. In global systolic function, the follow-up values of Sa glob in cohort I were improved by 0.23±0.36 as compared with baseline values at rest, and by 1.26±0.65 cm/s at the maximal load, while the values of Sa glob in cohort II were diminished by 0.53±0.22 (p=NS), and by 1.25±0.45 cm/s (p<0.05), respectively. Concerning the resting regional function, the only significant difference between cohorts in follow-up changes was found in walls E: 0.37±0.60 versus -1.76±0.40 cm/s (p<0.05). At the maximal load, the significant difference was found only in walls A (0.16±0.84 versus -2.67±0.87 cm/s; p<0.05). Patients with regular 12-month physical activity improved their global left ventricle systolic function mainly due to improvement of contractility in walls supplied by a totally occluded coronary artery.     

The [Ru(Hedta)NO](0.1-) system: structure, chemical reactivity and biological assays

The [Ru(II)(Hedta)NO(+)] complex is a diamagnetic species crystallizing in a distorted octahedral geometry, with the Ru-N(O) length 1.756(4) A and the RuNO angle 172.3(4) degrees . The complex contains one protonated carboxylate (pK(a)=2.7+/-0.1). The [Ru(II)(Hedta)NO(+)] complex undergoes a nitrosyl-centered one-electron reduction (chemical or electrochemical), with E(NO+/NO)=-0.31 V vs SCE (I=0.2 M, pH 1), yielding [Ru(II)(Hedta)NO](-), which aquates slowly: k(-NO)=2.1+/-0.4x10(-3) s(-1) (pH 1.0, I=0.2 M, CF(3)COOH/NaCF(3)COO, 25 degrees C). At pHs>12, the predominant species, [Ru(II)(edta)NO](-), reacts according to [Ru(II)(edta)NO](-)+2OH(-)-->[Ru(II)(edta)NO(2)](3-), with K(eq)=1.0+/-0.4 x 10(3) M(-2) (I=1.0 M, NaCl; T=25.0+/-0.1 degrees C). The rate-law is first order in each of the reactants for most reaction conditions, with k(OH(-))=4.35+/-0.02 M(-1)s(-1) (25.0 degrees C), assignable mechanistically to the elementary step comprising the attack of one OH(-) on [Ru(II)(edta)NO](-), with subsequent fast deprotonation of the [Ru(II)(edta)NO(2)H](2-) intermediate. The activation parameters were DeltaH(#)=60+/-1 kJ/mol, DeltaS(#)=-31+/-3 J/Kmol, consistent with a nucleophilic addition process between likely charged ions. In the toxicity up-and-down tests performed with Swiss mice, no death was observed in all the doses administered (3-9.08 x 10(-5) mol/kg). The biodistribution tests performed with Wistar male rats showed metal in the liver, kidney, urine and plasma. Eight hours after the injection no metal was detected in the samples. The vasodilator effect of [Ru(II)(edta)NO](-) was studied in aortic rings without endothelium, and was compared with sodium nitroprusside (SNP). The times of maximal effects of [Ru(II)(edta)NO](-) and SNP were 2 h and 12 min, respectively, suggesting that [Ru(II)(edta)NO](-) releases NO slowly to the medium in comparison with SNP.     

Comparison of blood pro/antioxidant levels before and after acute exercise in athletes and non-athletes

The aims of our study were to assess the redox state of adolescent athletes and non-athletes both at rest and after acute exposure to physical load and to find relations between parameters of redox state and morphofunctional characteristics of subjects. 58 young handball players and 37 non-athletes were subjected to body composition analysis, measuring of maximal oxygen consumption and blood sampling immediately before and after a maximal progressive exercise test. At rest, athletes had significantly higher superoxide dismutase (SOD) and catalase (CAT) activity, higher levels of glutathione (GSH) and nitric oxide (NO) and lower levels of lipid peroxidation (TBARS) compared with non-athletes. A maximal exercise test induced statistically significant rise of superoxide anion radical (O2-), hydrogen peroxide (H2O2) and NO levels in non-athletes, while TBARS levels decreased. Athletes experienced the fall in NO levels and the fall in CAT activity. After exercise, athletes had significantly lower levels of O2- compared with non-athletes. Two way repeated measures ANOVA showed that the response of O2-, NO and TBARS to the exercise test was dependent on the sports engagement (training experience) of subjects. Significant correlations between morphofunctional and redox parameters were found. These results suggest that physical fitness affects redox homeostasis.     

The influence of resting period length on jumping performance

The purpose of this study was to determine a resting interval between countermovement jumps (i.e., volleyball spikes) that allows the maintenance of maximal jumping performance. Ten male volleyball players (1.85 +/- 0.05 m, 77.2 +/- 10.6 kg, 21.6 +/- 5.3 years) performed 6 experimental jumping sessions. In the first and sixth sessions, maximal countermovement jump height was measured, followed by submaximal countermovement jumps to the point of volitional fatigue. The number of countermovement jumps was used as a reference to test the effect of rest period between volleyball spikes. From the second to fifth experimental sessions, 30 maximal volleyball spikes were performed with different resting periods (i.e., 8, 14, 17, and 20 seconds) followed by countermovement jumps. Between the 15th and 30th spikes, the blood lactate concentration and heart rate were measured. Because the performance on the first and sixth sessions was the same, no training effects were noticed. During the 8-second resting interval set, the lactate concentration increased significantly between the 15th and 30th spikes (i.e., from 3.37 +/- 1.16 mmol to 4.94 +/- 1.49 mmol); the number of countermovement jumps decreased significantly after spikes compared to those performed without a previous effort (i.e., from 23 +/- 7 jumps to 17 +/- 9 jumps); and these variables were significantly correlated (r = -0.7). On the other hand, the lactate concentration and number of countermovement jumps were stable across the other resting intervals, without a heart rate steady state. The results indicate that an adequate resting period between spikes allowed participants to achieve a lactate steady state in which the performance was maintained during the exercise. These findings show that resting intervals between 14 and 17 seconds, typical during volleyball matches, are indicated to use in volleyball spike drills due to their capacity to maintain maximal jumping performance.     

Effects of habitual smoking on cardiorespiratory responses to sub-maximal exercise

The effects of habitual cigarette smoking on cardiorespiratory responses to sub-maximal and maximal work were evaluated in nine adult nonsmokers and nine smokers with a mean age of 33 yr. A maximal treadmill test was followed by three tests at 45, 60 and 75% of each subject's VO(2)max. Compared to nonsmokers, the habitual smokers had a non-significantly lower VO(2)max in L/min and per lean body mass (9 and 6%, respectively), but had higher %fat (p<0.01), resulting in a significantly lower VO(2)max per kg body wt (13%, p<0.03). Maximal exercise ventilation (V(E)) was 16% lower in smokers. During sub-maximal work at equivalent exercise stress levels in the two groups, the V(E)/VO(2) ratio was higher in smokers by an average of 11% because VO(2) was lower and the respiratory exchange ratio values were significantly elevated in smokers at 75% of VO(2)max. Blood lactate concentrations in smokers were higher as workloads increased and O(2) pulse (VO(2)/HR) was significantly lower throughout, indicating reduced O(2) extraction, probably due to carbon monoxide. The resting HR was significantly higher in smokers and the HR recovery following all three submaximal exercises was significantly slower in smokers. These results show that detrimental cardiorespiratory effects of chronic cigarette smoking in apparently healthy individuals are evident at moderate exercise levels as reduced gas exchange efficiency in lungs and muscles.     

Dynamic exercise training in foxhounds. I. Oxygen consumption and hemodynamic responses

Ten foxhounds were studied during maximal and submaximal exercise on a motor-driven treadmill before and after 8-12 wk of training. Training consisted of working at 80% of maximal heart rate 1 h/day, 5 days/wk. Maximal O2 consumption (VO2max) increased 28% from 113.7 +/- 5.5 to 146.1 +/- 5.4 ml O2 X min-1 X kg-1, pre- to posttraining. This increase in VO2max was due primarily to a 27% increase in maximal cardiac output, since maximal arteriovenous O2 difference increased only 4% above pretraining values. Mean arterial pressure during maximal exercise did not change from pre- to posttraining, with the result that calculated systemic vascular resistance (SVR) decreased 20%. There were no training-induced changes in O2 consumption, cardiac output, arteriovenous O2 difference, mean arterial pressure, or SVR at any level of submaximal exercise. However, if post- and pretraining values are compared, heart rate was lower and stroke volume was greater at any level of submaximal exercise. Venous lactate concentrations during a given level of submaximal exercise were significantly lower during posttraining compared with pretraining, but venous lactate concentrations during maximal exercise did not change as a result of exercise training. These results indicate that a program of endurance training will produce a significant increase in VO2max in the foxhound. This increase in VO2max is similar to that reported previously for humans and rats but is derived primarily from central (stroke volume) changes rather than a combination of central and peripheral (O2 extraction) changes.     

Exercise training reduces PGE2 levels and induces recovery from steatosis in tumor-bearing rats

The effects of endurance training on PGE (2) levels and upon the maximal activity of hepatic carnitine palmitoyltransferase (CPT) system were studied in rats bearing the Walker 256 carciosarcoma. Animals were randomly assigned to a sedentary control (SC), sedentary tumor-bearing (ST), exercised control (EC), and as an exercised tumor-bearing (ET) group. Trained rats ran on a treadmill (60% VO (2) max) for 60 min/day, 5 days/week, for 8 weeks. We examined the mRNA expression (RT-PCR) and maximal activity (radioassay) of the carnitine palmitoyltransferase system enzymes (CPT I and CPT II), as well as the gene expression of fatty-acid-binding protein (L-FABP) in the liver. PGE (2) content was measured in the serum, in tumor cells, and in the liver (ELISA). CPT I and CPT II maximal activity were decreased (p<0.01) in ST when compared with SC. In contrast, serum PGE (2) was increased (p<0.05) in cachectic animals as compared with SC. In the liver, PGE (2) content was also increased (p<0.05) when compared with SC. Endurance training restored maximal CPT I and CPT II activity in the tumor-bearing animals (p<0.0001). Exercise training induced PGE (2) levels to return to control values in the liver of tumor-bearing training rats (p<0.05) and decreased the eicosanoid content in the tumor (p<0.01). In conclusion, endurance training was capable of reestablishing liver carnitine palmitoyltransferase (CPT) system activity associated with decreased PGE (2) levels in cachectic tumor-bearing animals, preventing steatosis.     

Cytotoxicity and site-specific DNA damage induced by nitroxyl anion (NO(-)) in the presence of hydrogen peroxide. Implications for various pathophysiological conditions.

Nitroxyl anion (NO(-)), the one-electron reduction product of nitric oxide (NO(.)), is formed under various physiological conditions. We have used four different assays (DNA strand breakage, 8-oxo-deoxyguanosine formation in calf thymus DNA, malondialdehyde generation from 2'-deoxyribose, and analysis of site-specific DNA damage using (32)P-5'-end-labeled DNA fragments of the human p53 tumor suppressor gene and the c-Ha-ras-1 protooncogene) to study the effects of NO(-) generated from Angeli's salt on DNA damage. It was found that strong oxidants are generated from NO(-), especially in the presence of H(2)O(2) plus Fe(III)-EDTA or Cu(II). NO(.) released from diethylamine-NONOate had no such effect. Distinct effects of hydroxyl radical (HO(.)) scavengers and patterns of site-specific DNA cleavage caused by Angeli's salt alone or by Angeli's salt, H(2)O(2) plus metal ion suggest that NO(-) acts as a reductant to catalyze the formation of the HO(.) from H(2)O(2) plus Fe(III) and formation of Cu(I)-peroxide complexes with a reactivity similar to HO(.) from H(2)O(2) and Cu(II). Angeli's salt and H(2)O(2) exerted synergistically cytotoxic effects to MCF-7 cells, determined by lactate dehydrogenase release assay. Thus NO(-) may play an important role in the etiology of various pathophysiological conditions such as inflammation and neurodegenerative diseases, especially when H(2)O(2) and transition metallic ions are present.     

Effects of high-intensity training on muscle lactate transporters and postexercise recovery of muscle lactate and hydrogen ions in women

The purpose of this study was to investigate the effects of high-intensity interval training (3 days/wk for 5 wk), provoking large changes in muscle lactate and pH, on changes in intracellular buffer capacity (betam(in vitro)), monocarboxylate transporters (MCTs), and the decrease in muscle lactate and hydrogen ions (H+) after exercise in women. Before and after training, biopsies of the vastus lateralis were obtained at rest and immediately after and 60 s after 45 s of exercise at 190% of maximal O2 uptake. Muscle samples were analyzed for ATP, phosphocreatine (PCr), lactate, and H+; MCT1 and MCT4 relative abundance and betam(in vitro) were also determined in resting muscle only. Training provoked a large decrease in postexercise muscle pH (pH 6.81). After training, there was a significant decrease in betam(in vitro) (-11%) and no significant change in relative abundance of MCT1 (96 +/- 12%) or MCT4 (120 +/- 21%). During the 60-s recovery after exercise, training was associated with no change in the decrease in muscle lactate, a significantly smaller decrease in muscle H+, and increased PCr resynthesis. These results suggest that increases in betam(in vitro) and MCT relative abundance are not linked to the degree of muscle lactate and H+ accumulation during training. Furthermore, training that is very intense may actually lead to decreases in betam(in vitro). The smaller postexercise decrease in muscle H+ after training is a further novel finding and suggests that training that results in a decrease in H+ accumulation and an increase in PCr resynthesis can actually reduce the decrease in muscle H+ during the recovery from supramaximal exercise.     

Detraining increases body fat and weight and decreases VO2peak and metabolic rate

Competitive collegiate swimmers commonly take a month off from swim training after their last major competition. This abrupt cessation of intense physical training has not been well studied and may lead to physiopsychological decline. The purpose of this investigation was to examine the effects of swim detraining (DT) on body composition, aerobic fitness, resting metabolism, mood state, and blood lipids in collegiate swimmers. Eight healthy endurance-trained swimmers (V(O2)peak, 46.7 ± 10.8 ml · kg(-1) · min(-1)) performed 2 identical test days, 1 in the trained (TR) state and 1 in the detrained (~5 weeks) state (DT). Body composition and circumferences, maximal oxygen consumption (V(O2)peak), resting metabolism (RMR), blood lipids, and mood state were measured. After DT, body weight (TR, 68.9 ± 9.7 vs. DT, 69.8 ± 9.8 kg; p = 0.03), fat mass (TR, 14.7 ± 7.6 vs. DT, 16.5 ± 7.4 kg; p = 0.001), and waist circumference (TR, 72.7 ± 3.1 vs. DT, 73.8 ± 3.6 cm; p = 0.03) increased, whereas V(O2)peak (TR, 46.7 ± 10.8 vs. DT, 43.1 ± 10.3 ml · kg(-1) · min(-1); p = 0.02) and RMR (TR, 1.34 ± 0.2 vs. DT, 1.25 ± 0.17 kcal · min(-1); p = 0.008) decreased, and plasma triglycerides showed a trend to increase (p = 0.065). Our data suggest that DT after a competitive collegiate swim season adversely affects body composition, fitness, and metabolism. Athletes and coaches need to be aware of the negative consequences of detraining from swimming, and plan off-season training schedules accordingly to allow for adequate rest/recovery and prevent overuse injuries. It's equally important to mitigate the negative effects on body composition, aerobic fitness and metabolism so performance may continue to improve over the long term.     

Effect of extracellular osmolality on cell volume and resting metabolism in mammalian skeletal muscle

The purpose of the present investigation was to establish an in vitro mammalian skeletal muscle model to study acute alterations in resting skeletal muscle cell volume. Isolated, whole muscles [soleus and extensor digitorum longus (EDL)] were dissected from Long-Evans rats and incubated for 60 min in Sigma medium 199 (1 g of resting tension, bubbled with 95% O(2)-5% O(2), 30 +/- 2 degrees C, and pH 7.4). Medium osmolality was altered to simulate hyposmotic (190 +/- 10 mmol/kg) or hyperosmotic conditions (400 +/- 10 mmol/kg), whereas an isosmotic condition (290 +/- 10 mmol/kg) served as a control. After incubation, relative water content of the muscle decreased with hyperosmotic and increased with hyposmotic condition in both muscle types (P < 0.05). The cross-sectional area of soleus type I and type II fibers increased (P < 0.05) in hyposmotic, whereas hyperosmotic exposure led to no detectable changes. The EDL type II fiber area decreased in the hyperosmotic condition and increased after hyposmotic exposure, whereas no change was observed in EDL type I fibers. Furthermore, exposure to the hyperosmotic condition in both muscle types resulted in decreased muscle ATP and phosphocreatine (P < 0.05) contents and increased creatine and lactate contents (P < 0.05) compared with control and hyposmotic conditions. This isolated skeletal muscle model proved viable and demonstrated that altering extracellular osmolality could cause acute alterations in muscle water content and resting muscle metabolism.     

Age-related augmentation of plasma catecholamines during dynamic exercise in healthy males

Although plasma norepinephrine (NE) increases with age in response to a variety of submaximal adrenergic stimuli, the effect of age on plasma catecholamine levels during maximal aerobic effort and during submaximal work at a fixed percent of peak O2 consumption (VO2) is unknown. We therefore measured NE, epinephrine (E), and VO2 at rest and during graded maximal treadmill exercise in 24 healthy male volunteers (ages 22-77 yr) from the Baltimore Longitudinal Study of Aging who were rigorously screened to exclude the presence of cardiovascular disease. At rest neither heart rate (HR) nor VO2 were age related. Resting NE (pg/ml) was not age related, but resting E (pg/ml) was higher in male subjects 68-77 yr old (group III) than in those aged 22-37 (group I) or 44-55 yr (group II), P less than 0.01. Maximal HR (beats/min) showed a strong inverse relationship to age (203.5 - 0.65 age, r = -0.80, P less than 0.001). Peak VO2 in milliliters per kilogram total body weight per minute decreased with age (47.7 - 0.23 age, r = -0.71, P less than 0.001). At maximal effort both NE (P less than 0.01) and E (P less than 0.05) were higher in group III than in either of the younger groups. At submaximal work levels NE and E also increased with age, and when normalized for relative effort at loads between 45 and 80% of peak VO2 both NE and E were higher in the group III male subjects, although statistical significance was reached for NE (P less than 0.01) but not for E (P = 0.09).(ABSTRACT TRUNCATED AT 250 WORDS)     

Hyperammoniemia during prolonged exercise: an effect of glycogen depletion?

Eight healthy men exercised to exhaustion on a cycle ergometer at a work load of 176 +/- 9 (SE) W corresponding to 67% (range 63-69%) of their maximal O2 uptake (exercise I). Exercise of the same work load was repeated after 75 min of recovery (exercise II). Exercise duration (range) was 65 (50-90) and 21 (14-30) min for exercise I and II, respectively. Femoral venous blood samples were obtained before and during exercise and analyzed for NH3 and lactate. Plasma NH3 was 12 +/- 2 and 19 +/- 6 mumol/l before exercise I and II, respectively and increased during exercise to exhaustion to peak values of 195 +/- 29 (exercise I) and 250 +/- 30 (exercise II) mumol/l, respectively. Plasma NH3 increased faster during exercise II compared with exercise I and at the end of exercise II was threefold higher than the value for the corresponding time of exercise I (P less than 0.001). Blood lactate increased during exercise I and after 20 min of exercise was 3.7 +/- 0.4 mmol/l and remained unchanged until exhaustion. During exercise II blood lactate increased less than during exercise I. It is concluded that long-term exercise to exhaustion results in large increases in plasma NH3 despite relatively low levels of blood lactate. It is suggested that the faster increase in plasma NH3 during exercise II (vs. exercise I) reflects an increased formation in the working muscle that may be caused by low glycogen levels and impairment of the ATP resynthesis.     

Training at high exercise intensity promotes qualitative adaptations of mitochondrial function in human skeletal muscle

This study explored mitochondrial capacities to oxidize carbohydrate and fatty acids and functional optimization of mitochondrial respiratory chain complexes in athletes who regularly train at high exercise intensity (ATH, n = 7) compared with sedentary (SED, n = 7). Peak O(2) uptake (Vo(2max)) was measured, and muscle biopsies of vastus lateralis were collected. Maximal O(2) uptake of saponin-skinned myofibers was evaluated with several metabolic substrates [glutamate-malate (V(GM)), pyruvate (V(Pyr)), palmitoyl carnitine (V(PC))], and the activity of the mitochondrial respiratory complexes II and IV were assessed using succinate (V(s)) and N,N,N',N'-tetramethyl-p-phenylenediamine dihydrochloride (V(TMPD)), respectively. Vo(2max) was higher in ATH than in SED (57.8 +/- 2.2 vs. 31.4 +/- 1.3 ml.min(-1).kg(-1), P < 0.001). V(GM) was higher in ATH than in SED (8.6 +/- 0.5 vs. 3.3 +/- 0.3 micromol O(2).min(-1).g dry wt(-1), P < 0.001). V(Pyr) was higher in ATH than in SED (8.7 +/- 1.0 vs. 5.5 +/- 0.2 micromol O(2).min(-1).g dry wt(-1), P < 0.05), whereas V(PC) was not significantly different (5.3 +/- 0.9 vs. 4.4 +/- 0.5 micromol O(2).min(-1).g dry wt(-1)). V(S) was higher in ATH than in SED (11.0 +/- 0.6 vs. 6.0 +/- 0.3 micromol O(2).min(-1).g dry wt(-1), P < 0.001), as well as V(TMPD) (20.1 +/- 1.0 vs. 16.2 +/- 3.4 micromol O(2).min(-1).g dry wt(-1), P < 0.05). The ratios V(S)/V(GM) (1.3 +/- 0.1 vs. 2.0 +/- 0.1, P < 0.001) and V(TMPD)/V(GM) (2.4 +/- 1.0 vs. 5.2 +/- 1.8, P < 0.01) were lower in ATH than in SED. In conclusion, comparison of ATH vs. SED subjects suggests that regular endurance training at high intensity promotes the enhancement of maximal mitochondrial capacities to oxidize carbohydrate rather than fatty acid and induce specific adaptations of the mitochondrial respiratory chain at the level of complex I.     

Fetal breathing and cardiovascular responses to graded methemoglobinemia in sheep

Graded methemoglobinemia (MetHb) was produced in unanesthetized fetal sheep to determine the effects on brain oxygenation. MetHb was induced by infusing methemoglobin-containing erythrocytes in exchange for fetal blood. During the hour after MetHb was established, fetal methemoglobin concentrations averaged 1.23 +/- 0.12 (mild MetHb), 1.71 +/- 0.13 (moderate MetHb), and 2.27 +/- 0.17 g/dl (severe MetHb). MetHb reduced mean arterial O2 content by approximately 19 (mild MetHb), 29 (moderate MetHb), and 39% (severe MetHb). The average preductal arterial PO2 fell by 1.6 (-7%), 2.8 (-11%), and 4.0 Torr (-16%) for mild, moderate, and severe MetHb, respectively. Fetal heart rate increased significantly during mild and moderate MetHb, and mean arterial pressure fell slightly during moderate and severe MetHb. The incidences of fetal breathing and eye movements were reduced in a dose-dependent manner when the calculated brain end-capillary PO2 was less than 14 Torr. We conclude that: 1) the effective capillary PO2 in the fetal brain can be significantly reduced by increasing the distance between non-methemoglobin-laden erythrocytes in capillaries and 2) hypoxic inhibition of fetal breathing probably arises from discrete areas of the brain having a PO2 less than 3 Torr.     

Relation between maximal aerobic power and the ability to repeat sprints in young basketball players

The aim of this study was to examine the effects of maximal aerobic power (V(.-)O2max peak) level on the ability to repeat sprints (calculated as performance decrement and total sprinting time) in young basketball players. Subjects were 18 junior, well-trained basketball players (age, 16.8 +/- 1.2 years; height, 181.3 +/- 5.7 cm; body mass, 73 +/- 10 kg; V(.-)O2max peak, 59.6 +/- 6.9 ml x kg(-1) x min(-1)). Match analysis and time-motion analysis of competitive basketball games was used to devise a basketball-specific repeated-sprint ability protocol consisting of ten 15-m shuttle run sprints with 30 s of passive recovery. Pre, post, and post plus 3-minute blood lactate concentrations were 2.5 +/- 0.7, 13.6 +/- 3.1, and 14.2 +/- 3.5 mmol x L(-1), respectively. The mean fatigue index (FI) value was 3.4 +/- 2.3% (range, 1.1-9.1%). No significant correlations were found between V(.-)O2max peak and either FI or total sprint time. A negative correlation (r = -0.75, p = 0.01) was found between first-sprint time and FI. The results of this study showed that V(.-)O2max peak is not a predictor of repeated-sprint ability in young basketball players. The high blood lactate concentrations found at the end of the repeated-sprint ability protocol suggest its use for building lactate tolerance in conditioned basketball players.     

Steady-state and transient kinetics of Escherichia coli nitric-oxide dioxygenase (flavohemoglobin). The B10 tyrosine hydroxyl is essential for dioxygen binding and catalysis

Escherichia coli expresses an inducible flavohemoglobin possessing robust NO dioxygenase activity. At 37 degrees C, the enzyme shows a maximal turnover number (V(max)) of 670 s(-1) and K(m) values for NADH, NO, and O(2) equal to 4.8, 0.28, and approximately 100 microM, respectively. Individual reduction, ligand binding, and NO dioxygenation reactions were examined at 20 degrees C, where V(max) is approximately 94 s(-1). Reduction by NADH occurs in two steps. NADH reduces bound FAD with a rate constant of approximately 15 microM(-1) s(-1), and heme iron is reduced by FADH(2) with a rate constant of 150 s(-1). Dioxygen binds tightly to reduced flavohemoglobin, with association and dissociation rate constants equal to 38 microM(-1) s(-1) and 0.44 s(-1), respectively, and the oxygenated flavohemoglobin dioxygenates NO to form nitrate. NO also binds reversibly to reduced flavohemoglobin in competition with O(2), dissociates slowly, and inhibits NO dioxygenase activity at [NO]/[O(2)] ratios of 1:100. Replacement of the heme pocket B10 tyrosine with phenylalanine increases the O(2) dissociation rate constant approximately 80-fold and reduces NO dioxygenase activity approximately 30-fold, demonstrating the importance of the tyrosine hydroxyl for O(2) affinity and NO scavenging activity. At 37 degrees C, V(max)/K(m)(NO) is 2,400 microM(-1) s(-1), demonstrating that the enzyme is extremely efficient at converting toxic NO into nitrate under physiological conditions.     

Nitric oxide, complex I, and the modulation of mitochondrial reactive species in biology and disease

Mitochondria are the specialized organelles for energy metabolism but also participate in the production of O(2) active species, cell cycle regulation, apoptosis and thermogenesis. Classically, regulation of mitochondrial energy functions was based on the ADP/ATP ratio, which dynamically stimulates the transition between resting and maximal O(2) uptake. However, in the last years, NO was identified as a physiologic regulator of electron transfer and ATP synthesis by inhibiting cytochrome oxidase. Additionally, NO stimulates the mitochondrial production of O(2) active species, primarily O(2)(-) and H(2)O(2), and, depending on NO matrix concentration, of ONOO(-), which is responsible for the nitrosylation and nitration of mitochondrial components. By this means, alteration in mitochondrial complexes restricts energy output, further increases O(2) active species and changes cell signaling for proliferation and apoptosis through redox effects on specific pathways. These mechanisms are prototypically operating in prevalent generalized diseases like sepsis with multiorgan failure or limited neurodegenerative disorders like Parkinson's disease. Complex I appears to be highly susceptible to ONOO(-) effects and nitration, which defines an acquired group of mitochondrial disorders, in addition to the genetically induced syndromes. Increase of mitochondrial NO may follow over-expression of nNOS, induction and translocation of iNOS, and activation and/or increased content of the newly described mtNOS. Likewise, mtNOS is important in the modulation of O(2) uptake and cell signaling, and in mitochondrial pathology, including the effects of aging, dystrophin deficiency, hypoxia, inflammation and cancer.