Transient oxygen uptake response to exercise characterizes functional capacity of the cardiocirculatory system in patients with chronic heart failure: a random stimulus approach

The transient response of oxygen uptake (V˙O₂) to submaximal exercise, known to be abnormal in patients with cardiovascular disorders, can be useful in assessing the functional status of the cardiocirculatory system, however, a method for evaluating it accurately has not yet been established. As an alternative approach to the conventional test at constant exercise intensity, we applied a random stimulus technique that has been shown to provide relatively noise immune responses of system being investigated. In 27 patients with heart failure and 24 age-matched control subjects, we imposed cycle exercise at 50 W intermittently according to a pseudo-random binary (exercise-rest) sequence, while measuring breath-by-breath V˙O₂. After determining the transfer function relating exercise intensity (W˙) to V˙O₂ and attenuating the high frequency ranges (>6 exercise-rest cycles · min⁻¹), we computed the high resolution band-limited (0–6 cycles · min⁻¹) V˙O₂ response (0–120 s) to a hypothetical step exercise. The V˙O₂ response showed a longer time constant in the patients than in the control subjects [47 (SD 37) and 31 (SD 8) s, respectively, P < 0.05]. Furthermore, the amplitude of the V˙O₂ response after the initial response was shown to be significantly smaller in the patients than in the control subjects [176 (SD 50) and 267 (SD 54) ml · min⁻¹ at 120 s]. The average amplitude over 120 s correlated well with peak V˙O₂ (r = 0.73) and ΔV˙O₂/ΔW˙ (r = 0.70), both of which are well-established indexes of exercise tolerance. The data indicated that our band-limited V˙O₂ step response using random exercise was more markedly attenuated and delayed in the patients with heart failure than in the normal controls and that it could be useful in quantifying the overall functional status of the cardiocirculatory system. Accepted: 6 January 1998     

Muscle oxygenation during incremental arm and leg exercise in men and women

The purpose of this study was to compare the rates of muscle deoxygenation in the exercising muscles during incremental arm cranking and leg cycling exercise in healthy men and women. Fifteen men and 10 women completed arm cranking and leg cycling tests to exhaustion in separate sessions in a counterbalanced order. Cardiorespiratory measurements were monitored using an automated metabolic cart interfaced with an electrocardiogram. Tissue absorbency was recorded continuously at 760 nm and 850 nm during incremental exercise and 6 min of recovery, with a near infrared spectrometer interfaced with a computer. Muscle oxygenation was calculated from the tissue absorbency measurements at 30%, 45%, 60%, 75% and 90% of peak oxygen uptake (V˙O₂) during each exercise mode and is expressed as a percentage of the maximal range observed during exercise and recovery (%Mox). Exponential regression analysis indicated significant inverse relationships (P < 0.01) between %Mox and absolute V˙O₂ during arm cranking and leg cycling in men (multiple R = −0.96 and −0.99, respectively) and women (R =−0.94 and −0.99, respectively). No significant interaction was observed for the %Mox between the two exercise modes and between the two genders. The rate of muscle deoxygenation per litre of V˙O₂ was 31.1% and 26.4% during arm cranking and leg cycling, respectively, in men, and 26.3% and 37.4% respectively, in women. It was concluded that the rate of decline in %Mox for a given increase in V˙O₂ between 30% and 90% of the peak V˙O₂ was independent of exercise mode and gender. Accepted: 31 March 1998     

The effect of a thiamin derivative on exercise performance

The purpose of this study was to investigate the effect of a thiamin derivative, thiamin tetrahydrofurfuryl disulfide (TTFD), on oxygen uptake (˙VO₂), lactate accumulation and cycling performance during exercise to exhaustion. Using a randomized, double-blind, cross-over design with a 10-day washout between trials, 14 subjects ingested either 1 g · day⁻¹ of TTFD or a placebo (PL) for 4 days. On day 3, subjects performed a progressive exercise test to exhaustion on a cycle ergometer for the determination of ˙VO_2submax, ˙VO_2peak, lactate concentration ([La⁻ ]), lactate threshold (Th_La) and heart rate ( f _c). On day 4, subjects performed a maximal 2000-m time trial on a cycle ergometer. A one-way analysis of variance (ANOVA) with repeated measures was used to determine significant differences between trials. There were no significant differences detected between trials for serial measures of ˙VO_2submax, [La⁻] or f _c. Likewise, ˙VO_2peak [PL 4.06 (0.19) TTFD 4.12 (0.19) l · min⁻¹, P = 0.83], Th_La [PL 2.47 (0.17), TTFD 2.43 (0.16) l · min⁻¹, P = 0.86] and 2000-m performance time [PL 204.5 (5.5), TTFD 200.9 (4.3) s, P = 0.61] were not significantly different between trials. The results of this study suggest that thiamin derivative supplementation does not influence high-intensity exercise performance. Accepted: 19 December 1996     

Noninvasive skeletal muscle lactate detection between periods of intense exercise in humans

We investigated whether localized ¹H nuclear magnetic resonance spectroscopy (NMRS) using stimulated echoes (STEAM) with a long mixing time (t _m) allowed the suppression of the fat signal and detection of lactate in skeletal muscle. The ¹H NMRS sequence was first validated in three isolated and perfused rabbit biceps brachii muscles. Spectra were obtained on a wide-bore spectrometer using a dual-tuned probe (¹H and ³¹P). Death was simulated by ceasing the muscle perfusion, which allowed post-mortem changes to be followed. During and after the simulated death, changes in levels of pH and in content of energy-rich compounds were observed with ³¹P NMRS. Our results showed an inverse linear relationship between pH and lactate in each of the three rabbits (r = 0.93, P < 0.001; r = 0.92, P < 0.01; r = 0.89, P < 0.01) and a decrease in phosphocreatine and concomitant increase in lactate. We then investigated whether this sequence allowed repeated detection of lactate in human soleus muscle during the recovery between periods of intense exercise (force-velocity test, F-v test). Seven subjects mean age 25.1 (SEM 0.8) years participated in this study. Soleus muscle lactate was detected at rest and for 3 min 30 s of the 5-min recovery between periods using a 2.35-T 40-cm bore magnet spectrometer. Arm venous plasma lactate concentration was measured at rest, during the F-v test when the subject stopped pedalling (S₁), and at the end of each 5-min recovery between periods (S₂). Results showed that the venous plasma lactate concentration at S₁ and S₂ increased significantly from the beginning of the F-v test to peak anaerobic power (W _an,peak) (P < 0.001). The spectra showed that muscle lactate resonance intensity rose markedly when W _an,peak was achieved. The muscle lactate resonance intensity plotted as a percentage of the resting value increased significantly at W _an,peak compared with submaximal braking forces (P < 0.05). We concluded from these results that localized ¹H NMRS using STEAM with a long t _m allows suppression of the fat signal and repeated detection of lactate on isolated perfused skeletal muscle in animals and between periods of intense exercise in humans. Accepted: 19 January 1998     

Anaerobic contribution to the time to exhaustion at the minimal exercise intensity at which maximal oxygen uptake occurs in elite cyclists, kayakists and swimmers

Using 23 elite male athletes (8 cyclists, 7 kayakists, and 8 swimmers), the contribution of the anaerobic energy system to the time to exhaustion (t _lim) at the minimal exercise intensity (speed or power) at which maximal oxygen uptake (V˙O_{2 max}) occurs (I _{V˙O2 max}) was assessed by analysing the relationship between the t _lim and the accumulated oxygen deficit (AOD). After 10-min warming up at 60% of V˙O_{2 max}, the exercise intensity was increased so that each subject reached his I _V˙O2max in 30 s and then continued at that level until he was exhausted. Pre-tests included a continuous incremental test with 2 min steps for determining the I _V˙O2max and a series of 5-min submaximal intensities to collect the data that would allow the estimation of the energy expenditure at I _V˙O2max . The AOD for the t _lim exercise was calculated as the difference between the above estimation and the accumulated oxygen uptake. The mean percentage value of energy expenditure covered by anaerobic metabolism was 15.2 [(SD 6)%, range 8.9–24.1] with significant differences between swimmers and kayakists (16.8% vs 11.5%, P≤0.05) and cyclists and kayakists (16.4% vs 11.5%, P≤0.05). Absolute AOD values ranged from 26.4 ml · kg⁻¹ to 83.6 ml · kg⁻¹ with a mean value of 45.9 (SD 18) ml · kg⁻¹. Considering all the subjects, the t _lim was found to have a positive and significant correlation with AOD (r = 0.62, P≤0.05), and a negative and significant correlation with V˙O_{2 max} (r = −0.46, P≤0.05). The data would suggest that the contribution of anaerobic processes during exercise performed at I _V˙O2max should not be ignored when t _lim is used as a supplementary parameter to evaluate specific adaptation of athletes. Accepted: 17 December 1996     

Water influx and efflux in free-flying pigeons

We used tritium-labeled water to measure total body water, water influx (which approximated oxidative water production) and water efflux in free-flying tippler pigeons (Columba livia) during flights that lasted on average 4.2 h. At experimental air temperatures ranging from 18 to 27 °C, mean water efflux by evaporation and excretion [6.3 ± 1.3 (SD) ml · h⁻¹, n = 14] exceeded water influx from oxidative water and inspired air (1.4 ± 0.7 ml · h⁻¹, n = 14), and the birds dehydrated at 4.9 ± 0.9 ml · h⁻¹. This was not significantly different from gravimetrically measured mass loss of 6.2 ± 2.1 g · h⁻¹ (t = 1.902, n = 14, P>0.05). This flight-induced dehydration resulted in an increase in plasma osmolality of 4.3 ± 3.0 mosmol · kg⁻¹ · h⁻¹ during flights of 3–4 h. At 27 °C, the increase in plasma osmolality above pre-flight levels (ΔP _osm = 7.6±4.29 mosmol · kg⁻¹ · h⁻¹, n = 6) was significantly higher than that at 18 °C (ΔP _osm = 0.83±2.23 mosmol · kg⁻¹ · h⁻¹, (t = 3.43, n = 6, P < 0.05). Post-flight haematocrit values were on average 1.1% lower than pre-flight levels, suggesting plasma expansion. Water efflux values during free flight were within 9% of those in the one published field study (Gessaman et al. 1991), and within the range of values for net water loss determined from mass balance during wind tunnel experiments (Biesel and Nachtigall 1987). Our net water loss rates were substantially higher than those estimated by a simulation model (Carmi et al. 1992) suggesting some re-evaluation of the model assumptions is required. Accepted: 8 April 1997     

Exercise-induced oxyhemoglobin desaturation and pulmonary diffusing capacity during high-intensity exercise

The purpose of this investigation was to examine if exercise-induced arterial oxyhemoglobin desaturation selectively observed in highly trained endurance athletes could be related to differences in the pulmonary diffusing capacity (D _L) measured during exercise. The D _L of 24 male endurance athletes was measured using a 3-s breath-hold carbon monoxide procedure (to give D _LCO) at rest as well as during cycling at 60% and 90% of these previously determined O_2max. Oxyhemoglobin saturation (S _aO₂%) was monitored throughout both exercise protocols using an Ohmeda Biox II oximeter. Exercise-induced oxyhemoglobin desaturation (DS) (S _aO₂% < 91% at O_2max) was observed in 13 subjects [88.2 (0.6)%] but not in the other 11 nondesaturation subjects [NDS: 92.9 (0.4)%] (P ≤ 0.05), although O_2max was not significantly different between the groups [DS: 4.34 (0.65) l / min vs NDS: 4.1 (0.49) l / min]. At rest, no differences in either D _LCO [m1 CO · mmHg⁻¹ · min⁻¹: 41.7 (1.7) (DS) vs 41.1 (1.8) (NDS)], D _LCO / _A [8.2 (0.4) (DS) vs 7.3 (0.9) (NDS)], MVV [l / min: 196.0 (10.4) (DS) vs 182.0 (9.9) (NDS)] or FEV₁/FVC [86.3 (2.2) (DS) vs 82.9 (4.7) (NDS)] were found between groups (P ≥ 0.05). However, _E /O₂ at O_2max was lower in the DS group [33.0 (1.1)] compared to the NDS group [36.8 (1.5)] (P ≤ 0.05). Exercise D _LCO (m1 CO · mmHg⁻¹ · min⁻¹) was not different between groups at either 60% O_2max [DS: 55.1 (1.4) vs NDS: 57.2 (2.1)] or at 90% O_2max [DS: 61.0 (1.8) vs NDS: 61.4 (2.9)]. A significant relationship (r = 0.698) was calculated to occur between S _aO₂% and _E /O₂ during maximal exercise. The present findings indicate that the exercise-induced oxyhemoglobin desaturation seen during submaximal and near-maximal exercise is not related to differences in D _L, although during maximal exercise S _aO₂ may be limited by a relatively lower exercise ventilation. Accepted: 25 September 1996     

Ventilatory efficiency and exercise tolerance in 101 healthy volunteers

The ventilatory equivalent for CO₂ defines ventilatory efficiency largely independent of metabolism. An impairment of ventilatory efficiency may be caused by an increase in either anatomical or physiological dead space, the latter being the most important mechanism in the hyperpnoea of heart failure, pulmonary embolism, pulmonary hypertension and the former in restrictive lung disease. However, normal values for ventilatory efficiency have not yet been established. We investigated 101 (56 men) healthy volunteers, aged 16–75 years, measuring ventilation and gas exchange at rest (n = 64) and on exercise (modified Naughton protocol, n = 101). Age and sex dependent normal values for ventilatory efficiency at rest defined as the ratio ventilation:carbon dioxide output (V˙ _E:V˙CO₂), exercise ventilatory efficiency during exercise, defined as the slope of the linear relationship between ventilation and carbon dioxide output (V˙ _E vs V˙CO₂ slope), oxygen uptake at the anaerobic threshold and at maximum (V˙O_2AT,V˙O_2max, respectively) and breathing reserve were established. Ventilatory efficiency at rest was largely independent of age, but was smaller in the men than in the women [V˙ _E:V˙CO₂ 50.5 (SD 8.8) vs 57.6 (SD 12.6) P<0.05]. Ventilatory efficiency during exercise declined significantly with age and was smaller in the men than in the women (men: (V˙ _E vs V˙CO₂ slope = 0.13 × age + 19.9; women: V˙ _E vs V˙CO₂ slope = 0.12 × age + 24.4). The V˙O_2AT and V˙O_2max were 23 (SD 5) and 39 (SD 7) ml O₂ · kg · min⁻¹ in the men and 18 (SD 4) and 32 (SD 7) in the women, respectively, and declined significantly with age. The V˙O_2AT was reached at 58 (SD 9)% V˙O_2max. Breathing reserve at the end of exercise was 41% and was independent of sex and age. It was concluded from this study that ventilatory efficiency as well as peak oxygen uptake are age and sex dependent in adults. Accepted: 11 June 1997     

Effects of artificially-induced anaemia on sudomotor and cutaneous blood flow responses to heat stress

The influence of artificially induced anaemia on thermal strain was evaluated in trained males. Heat stress trials (38.6°C, water vapour pressure 2.74 kPa) performed at the same absolute work rates [20 min of seated rest, 20 min of cycling at 30% peak aerobic power (V˙O_2peak), and 20 min cycling at 45% V˙O_2peak] were completed before (HST1) and 3–5 days after 3 units of whole blood were withdrawn (HST2). Mild anaemia did not elevate thermal strain between trials, with auditory canal temperatures terminating at 38.5°C [(0.16), HST1] and 38.6°C [(0.13), HST2; P > 0.05]. Given that blood withdrawal reduced aerobic power by 16%, this observation deviates from the close association often observed between core temperature and relative exercise intensity. During HST2, the absolute and integrated forearm sweat rate (m˙ _sw) exceeded control levels during exercise (P < 0.05), while a suppression of forehead m˙ _sw occurred (P < 0.05). These observations are consistent with a possible peripheral redistribution of sweat secretion. It was concluded that this level of artificially induced anaemia did not impact upon heat strain during a 60-min heat stress test. Accepted: 17 April 1997     

Habitual physical activity and peak anaerobic power in elderly women

The aim of this study was to investigate the relationship between maximal anaerobic power (P _max) and corresponding optimal velocity (V _opt) and habitual physical activity (PA) on the one hand and with maximal oxygen consumption (V˙O_2max) on the other hand, in elderly women. Twenty-nine community dwelling, healthy women aged 66–82 years participated in the study. PA was evaluated using the Questionnaire d'Activite Physique Saint-Etienne (QAPSE) and expressed using two QAPSE activity indices: mean habitual daily energy expenditure (MHDEE) and daily energy expenditure corresponding to leisure time sports activities (sports activity). The subjects' P _max and V _opt were measured while they cycled on a friction-loaded non-isokinetic cycle ergometer. P _max was expressed relative to body mass [P _max/kg(W · kg⁻¹)], and relative to the mass of two quadriceps muscles [P _{max /Quadr}(W·kg_Quadr ⁻¹)]. A negative relationship between P _max/kg (Spearman's r = −0.56; P < 0.01), P _max/Quadr (r = −0.53; P < 0.01) and V _opt (r = −0.45; P < 0.05) and age was found. P _max/kg was positively associated with MHDEE (r = 0.51; P < 0.01) and sports activity (r = 0.58; P < 0.01), as were P _max/Quadr and V _opt (r = 0.55; P < 0.01 and r = 0.54; P < 0.01, respectively). P _max/kg, P _max/Quadr and V _opt correlated positively with V˙O_2max. The positive relationship between ergometer measurements and PA indices was similar to that between V˙O_2max and PA. P _max/kg was, moreover, closely related to V _opt (r = 0.77; P < 0.001). When a multiple stepwise regression analysis was used to select the variables influencing ergometer measurements, MHDEE contributed significantly to P _max/kg variance, whereas sports activity contributed to P _max/Quadr and V _opt variances. In conclusion, the data from this cross-sectional study suggest that in healthy elderly women habitual PA, and especially leisure time PA, alleviates the decline of the P _max of the quadriceps muscles. Accepted: 30 January 1997     

Kinetic studies on the reduction of the R2 subunit of mouse ribonucleotide reductase with hydroxyurea, hydrazine, phenylhydrazine and hydroxylamine

 Four reductions of the R2 subunit of mouse ribonucleotide reductase have been studied and found to exhibit different behaviour from that of Escherichia coli R2. An important difference is that there is no stable met-R2 (Fe₂ ^II I) form of mouse R2. With hydroxyurea, hydrazine and hydroxylamine uniphasic kinetics are observed for the combined reduction of radical Tyr ^˙ and Fe₂ ^II I components to tyrosine and Fe₂ ^II respectively. The rate constants, determined at 370 nm (emphasising Fe^III decay) and 417 nm (emphasising Tyr ^˙ decay), differ by factors of 2–3, allowing some mechanistic features to be defined. The studies with hydrazine are particularly important. In the case of E. coli R2, a first phase corresponding to two-equivalent reduction of the met-R2 component has been observed [18]. It is likely that the four times slower second phase reaction of active E. coli R2 also corresponds to the Fe₂ ^II I → Fe₂ ^II change and is followed by fast intramolecular Fe₂ ^II reduction of the higher potential Tyr ^˙. The latter changes are believed to hold also for (active) mouse R2. The Fe^IIFe^III semi forms have been detected at low levels by EPR for mouse R2 (9%) and E. coli (∼5%) in previous studies. Further substrate reduction of Fe^IIFe^III occurs at a comparable rate to account for the transient behaviour of Fe^IIFe^III. For mouse R2 the combined Fe^III decay processes (which we are unable to separate) give smaller uniphasic rate constants at 370 nm than at 417 nm. A fitted-base-line (FBL) treatment of absorbance changes at 417 nm targets more closely the Tyr ^˙ decay as a means of monitoring the rate-determining step. The FBL method gives rate constants k (M^–1 s^–1) at 25  °C and pH 7.5 for hydroxyurea (1.46), hydrazine (0.163) and hydroxylamine (4.4). Surprisingly, phenylhydrazine, with a less strong reduction potential (0.25 V), gives a substantially faster reduction of the Tyr ^˙ as the only redox step (rate constant 27 M^–1 s^–1). In this case a slower second phase at 370 nm is independent of reductant and corresponds to rate-controlling release of Fe^III. Overall the results indicate a more reactive redox centre for mouse R2 and help develop further an understanding of factors affecting the reactivity of R2. Received: 11 October 1996 / Accepted: 11 February 1997     

Running economy deteriorates following 60?min of exercise at 80% V˙O2max

Fifteen young adult Singaporean male physical education students maximum oxygen consumption [(V˙O_2max) = 56 (4.7) ml · kg⁻¹ · min⁻¹] performed three prolonged runs in a counterbalanced design. The running bouts varied in time (40 vs 60 min) and intensity (70% vs 80% V˙O_{2 max} ). Each prolonged run was separated by 7 days. The running economy (RE) at 10.8 km · h⁻¹ during 10-min running bouts was measured before (RE1) and after (RE2) each prolonged run. A control study involved monitoring RE at 10.8 km · h⁻¹ before and after 60 min rest. There were no differences between RE1 and RE2 values during the control run. However, there were differences between RE1 and RE2 values when separated by a prolonged run. For example, the mean (SD) changes in oxygen consumption (ml · kg⁻¹ · min⁻¹) values were 38.2 (2.5) versus 40.1 (2.6) (40 min at 80% V˙O_{2 max} ), 38.9 (2.8) versus 41.5 (2.6) (60 min at 70% V˙O_{2 max} ), and 39.0 (3.1) versus 42.7 (2.9) (60 min at 80% V˙O_{2 max} ; P < 0.01). The results of this investigation support the hypothesis that RE deteriorates during prolonged running, and that the magnitude of the deterioration in RE increases with both increasing exercise intensity and duration. Accepted: 14 July 1997     

Effects of amount of training on the saliva concentrations of cortisol, dehydroepiandrosterone and on the dehydroepiandrosterone: cortisol concentration ratio in women over 16 weeks of training

This study was designed to investigate in the saliva the influence in female athletes of handball or volleyball training on concentrations of cortisol [C], dehydroepiandrosterone [DHEA], and on the [DHEA]:[C] ratio over 16 weeks of training. Data were compared to those of sedentary women. Saliva samples were collected upon waking after an overnight fast during the 1st week (W₁) of the training programme and in the 16th week (W₁₆). The training programme increased the resting concentrations of saliva [DHEA] in all the sportswomen. In contrast, a decrease of [DHEA] was noted in the sedentary group (W₁₆ < W₁; P < 0.05). In none of the women did the [C] at rest change significantly during the study. Between W₁ and W₁₆, the [DHEA]:[C] ratio increased by more than 30% in all the sportswomen. In addition, the athletes with the highest performance levels and greatest amount of training had the lowest [DHEA]:[C] ratio. Negative linear relationships between the amount of training and the [DHEA]:[C] ratio were found both at W₁ (r = −0.53 P < 0.001), and W₁₆ (r=−0.73 P < 0.001), suggesting that the latter could be used as an indicator of the training status of sportswomen. Accepted: 12 May 1998     

Effects of trans-pyridine ligands on the interactions of RuII and RuIII ammine complexes N7-coordinated to purine nucleosides and DNA

 DNA binding by trans-[(H₂O)(Pyr)(NH₃)₄Ru^II]²⁺ (Pyr=py, 3-phpy, 4-phpy, 3-bnpy, 4-bnpy) is highly selective for G⁷ with K _G=1.1×10⁴ to 2.8×10⁴, with the more hydrophobic Pyr ligands exhibiting slightly higher binding. A strong dependence on ionic strength indicates that ion-pairing with DNA occurs prior to binding. At μ=0.05, d[Ru^II-DNA]/dt=k[Ru^II][DNA], where k=0.17–0.21 M^–1s^–1 with the various Pyr ligands. The air oxidation of [(py)(NH₃)₄Ru^II] _n -DNA to [(py)(NH₃)₄Ru^III] _n -DNA at pH 6 occurs with a pseudo-first-order rate constant of k _obs=5.6×10^–4s^–1 at μ=0.1, T=25  °C. Strand cleavage of plasmid DNA appears to occur by both Fenton/Haber-Weiss chemistry and by base-catalyzed routes, some of which are independent of oxygen. Base-catalyzed cleavage is more efficient than O₂ activation at neutral pH and involves the disproportionation of covalently bound Ru^III and, in the presence of O₂, Ru-facilitated autoxidation to 8-oxoguanine. Disproportionation of [py(NH₃)₄Ru^III] _n -DNA occurs according to the rate law: d[Ru^II–G_DNA]/dt=k ₀[Ru^III–G_DNA]+k ₁[Ru^III–G_DNA][OH^–], where k ₀=5.4×10^–4s^–1 and k ₁=8.8 M^–1s^–1 at 25  °C, μ=0.1. The appearance of [(Gua)(py)(NH₃)₄Ru^III] under argon, which occurs according to the rate law: d[Ru^III–G]/dt=k ₀[Ru^III–G_DNA]+k ₁[OH^–][Ru^III–G_DNA] (k ₀=5.74×10^–5 s^–1, k ₁=1.93×10^–2M^–1s^–1 at T=25  °C, μ=0.1), is consistent with lysis of the N-glycosidic bond by Ru^IV-induced general acid hydrolysis. In air, the ratio of [Ru-8-OG]/[Ru-G] and their net rates of appearance are 1.7 at pH 11, 25  °C. Small amounts of phosphate glycolate indicate a minor oxidative pathway involving C4′ of the sugar. In air, a dynamic steady-state system arises in which reduction of Ru^IV produces additional Ru^II. Received: 11 November 1998 / Accepted: 3 March 1999     

Energetics of swimming at maximal speeds in humans

The energy cost per unit of distance (C _s, kilojoules per metre) of the front-crawl, back, breast and butterfly strokes was assessed in 20 elite swimmers. At sub-maximal speeds (v), C _s was measured dividing steady-state oxygen consumption (V˙O₂) by the speed (v, metres per second). At supra-maximal v, C _s was calculated by dividing the total metabolic energy (E, kilojoules) spent in covering 45.7, 91.4 and 182.9 m by the distance. E was obtained as: E = E _an+V˙O_2max t _p−V˙O_2max(1−e^{−( t p/)}), where E _an was the amount of energy (kilojoules) derived from anaerobic sources, V˙O_2max litres per second was the maximal oxygen uptake, α (=20.9 kJ · l O₂ ⁻¹) was the energy equivalent of O₂, τ (24 s) was the time constant assumed for the attainment of V˙O_2max at muscle level at the onset of exercise, and t _p (seconds) was the performance time. The lactic acid component was assumed to increase exponentially with t _p to an asymptotic value of 0.418 kJ · kg⁻¹ of body mass for t _p ≥ 120 s. The lactic acid component of E _an was obtained from the net increase of lactate concentration after exercise (Δ[La]_b) assuming that, when Δ[La]_b = 1 mmol · l⁻¹ the net amount of metabolic energy released by lactate formation was 0.069 kJ · kg⁻¹. Over the entire range of v, front crawl was the least costly stroke. For example at 1 m · s⁻¹, C _s amounted, on average, to 0.70, 0.84, 0.82 and 0.124 kJ · m⁻¹ in front crawl, backstroke, butterfly and breaststroke, respectively; at 1.5 m · s⁻¹, C _s was 1.23, 1.47, 1.55 and 1.87 kJ · m⁻¹ in the four strokes, respectively. The C _s was a continuous function of the speed in all of the four strokes. It increased exponentially in crawl and backstroke, whereas in butterfly C _s attained a minimum at the two lowest v to increase exponentially at higher v. The C _s in breaststroke was a linear function of the v, probably because of the considerable amount of energy spent in this stroke for accelerating the body during the pushing phase so as to compensate for the loss of v occurring in the non-propulsive phase. Accepted: 14 April 1998     

The influence of either no fluid or carbohydrate-electrolyte fluid ingestion and the environment (thermoneutral versus hot and humid) on running economy after prolonged, high-intensity exercise

This study investigated the effects on running economy (RE) of ingesting either no fluid or an electrolyte solution with or without 6% carbohydrate (counterbalanced design) during 60-min running bouts at 80% maximal oxygen consumption (V˙O_2max). Tests were undertaken in either a thermoneutral (22–23°C; 56–62% relative humidity, RH) or a hot and humid natural environment (Singapore: 25–35°C; 66–77% RH). The subjects were 15 young adult male Singaporeans [V˙O_2max = 55.5 (4.4 SD) ml kg⁻¹ min⁻¹]. The RE was measured at 3 m s⁻¹ [65 (6)% V˙O_2max] before (RE1) and after each prolonged run (RE2). Fluids were administered every 2 min, at an individual rate determined from prior tests, to maintain body mass (group mean = 17.4 ml min⁻¹). The V˙O₂ during RE2 was higher (P < 0.05) than that during the RE1 test for all treatments, with no differences between treatments (ANOVA). The mean increase in V˙O₂ from RE1 to RE2 ranged from 3.4 to 4.7 ml kg⁻¹ min⁻¹ across treatments. In conclusion, the deterioration in RE at 3 m s⁻¹ (65% V˙O_2max) after 60 min of running at 80% V˙O_2max appears to occur independently of whether fluid is ingested and regardless of whether the fluid contains carbohydrates or electrolytes, in both a thermoneutral and in a hot, humid environment. Accepted: 30 October 1997     

Energetic cost of hovering flight in a nectar-feeding bat measured with fast-response respirometry

Hover-feeding glossophagine bats provide, in addition to the hummingbirds, a second vertebrate model for the analysis of hovering flight based on metabolic measurement and aerodynamic theory. In this study, the power input of hovering Glossophaga soricina bats (11.9 g) was measured by standard respirometry and fast-response (<0.2 s) oxygen analysis. Bats needed 5–7 s after a rest-to-flight transition to return to a respiratory steady state. Therefore, only hovering events preceeded by a 7-s flight interval were evaluated. V˙_O2 during hovering fluctuated with a frequency of 3–5 Hz, which corresponded in frequency to the licking movement of the tongue. During hovering, bats often may have hypoventilated as indicated by reduced V˙_O2 and a respiratory exchange ratio (RER) well below the steady-state value of 1. Steady-state oxygen consumption (and derived power input) during hovering was estimated to be 27 (25–29) ml O₂ g⁻¹ h⁻¹ (158 W kg⁻¹ or 1.88 W) in the 11.9-g bats as indicated by three independent findings: (1) V˙_O2 was 26 ml O₂ g⁻¹ h⁻¹ after 6.5 s of hovering, (2) the mean RER during single hovering events was at its steady-state level of 1 only at oxygen uptake rates of 25–29 ml g⁻¹ h⁻¹, and (3) when the oxygen potentially released from estimated oxygen stores was added to the measured oxygen uptake, the upper limit for oxygen consumption during hovering was found to be 29 ml O₂ g⁻¹ h⁻¹. Hovering power input was about 1.2 times the value of minimum flight power input (Winter and von Helversen 1998) and thus well below the 1.7–2.6 difference in power output postulated by aerodynamic theory (Norberg et al. 1993). Mass specific power input was 40% less than in hummingbirds. Thus, within the possible modes of hovering flight, Glossophaga bats seem to operate at the high-efficiency end of the spectrum. Accepted: 28 April 1998     

Assessment of middle-distance running performance in sub-elite young runners using energy cost of running

The purpose of this study was to assess the validity of v _amax as an indicator of middle-distance running performance in sub-elite young runners, _amax being defined as the quotient maximal oxygen uptake (V˙O _2max) divided by the net energy cost of running (C _r) on a treadmill at a submaximal running velocity (280 m · min⁻¹). The V˙O _2max, ventilatory threshold, _amax, and C _r were assessed in 39 young male sub-elite runners having only small variations in performance level. The relationship between each variable and running performance (at 1500 m, 3000 m, and 5000 m) was evaluated. A trend toward a negative correlation existed between C _r and performance although this was not significant. The V˙O _2max and _amax were significantly related to performance. The _amax accounted for around 50% of the variability in performance whereas other physiological variables selected in this study were responsible, at best, for approximately 39%. The results presented in this study suggested that _amax was a useful indicator of middle-distance running performance in sub-elite young runners with similar performance levels as well as in top elite athletes. Accepted: 19 August 1997     

Ion channels in guard cells of Arabidopsis thaliana (L.) Heynh.

Despite the availability of many mutants for signal transduction, Arabidopsis thaliana guard cells have so far not been used in electrophysiological research. Problems with the isolation of epidermal strips and the small size of A. thaliana guard cells were often prohibiting. In the present study these difficulties were overcome and guard cells were impaled with double-barreled microelectrodes. Membrane-potential recordings were often stable for over half an hour and voltage-clamp measurements could be conducted. The guard cells were found to exhibit two states. The majority of the guard cells had depolarized membrane potentials, which were largely dependent on external K⁺ concentrations. Other cells displayed spontaneous transitions to a more hyperpolarized state, at which the free-running membrane potential (E_m) was not sensitive to the external K⁺ concentration. Two outward-rectifying conductances were identified in cells in the depolarized state. A slow outward-rectifying channel (s-ORC) had properties resembling the K⁺-selective ORC of Vicia faba guard cells (Blatt, 1988, J Membr Biol 102: 235–246). The activation and inactivation times and the activation potential, all depended on the reversal potential (E_rev) of the s-ORC conductance. The s-ORC was blocked by Ba²⁺ (K_1/2 = 0.3–1.3mM) and verapamil (K_1/2 = 15–20 μM). A second rapid outward-rectifying conductance (r-ORC) activated instantaneously upon stepping the voltage to positive values and was stimulated by Ba²⁺. Inward-rectifying channels (IRC) were only observed in cells in the hyperpolarized state. The activation time and activation potential of this channel were not sensitive to the external K⁺ concentration. The slow activation of the IRC (t_1/2 ≈ 0.5 s) and its negative activation potential (V_threshold = −155 mV) resemble the values found for the KAT1 channel expressed in Saccharomyces cerevisiae (Bertl et al., 1995, Proc Natl Acad Sci USA 92: 2701–2705). The results indicate that A. thaliana guard cells provide an excellent system for the study of signal transduction processes. Received: 28 March 1996 / Accepted: 11 November 1996